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gen_d731ba179338909192f102e915432bd0 | Identifying strategies to reveal genetic results over the lifespan | NIH | BRIGHAM AND WOMEN'S HOSPITAL | 5K99HG012809-02 | PROJECT SUMMARY/ABSTRACT The widespread sequencing of healthy babies is imminent: at least a dozen research projects have recently launched, and several companies offer newborn genetic screening panels. A newborn’s genome can contain health information of relevance across their lifespan — as a baby, later in childhood, and in adulthood. This poses a timing issue: if babies are sequenced near birth, when should this information be revealed? A proposed vision for the future of genomic medicine is to reveal information as it becomes relevant, to the child’s parents and later, if desired, to the individual. This would necessitate the genome being kept “on file,” to be used as a resource over time. This strategy may promote the ethical rollout of lifelong genomic medicine by promoting the developing child’s autonomy and optimizing the balance of benefits to risks. However, the feasibility of this strategy, the details of its implementation, and its implications have yet to be explored in a rigorous and empirical manner. Perhaps other approaches are preferable. A second, simplified, strategy would reveal all childhood-relevant information at birth and then give the individual the option of receiving adult-onset information at age 18. A third strategy would reject using the genome as a resource over time, and just generate one report for a baby, potentially including adult-onset information. This strategy may be preferable because the use of the genome as a resource raises complex ethical, legal, and social implications (ELSI), including data control, privacy, consent, legal obligations, and decision making about when information becomes relevant. For these different strategies, this project will 1) Determine their feasibility, 2) Assess their ELSI, 3) Understand the preferences of parents from diverse backgrounds, and 4) Develop consensus on the necessary and desirable features for a strategy to sequence babies near birth, possibly using the genome as a resource over time. The project will have impact by producing concrete, evidence-based and ethically framed recommendations for implementers of newborn sequencing. The candidate was originally trained as a computational biologist, was formerly employed in the genomics industry, and is currently an ELSI scholar. Her goal is to become an independent investigator working in the context of clinical research informing the adoption of genomic medicine to identify, assess and address ELSI questions, ultimately to ensure that genomic medicine works to the benefit of all. To accomplish this goal, this proposal focuses her training efforts on a) developing skills in conducting surveys, b) developing expertise in Delphi methods, and c) refining skills in conceptual and normative analysis. The project will leverage the BabySeq cohorts, the world’s first empirical studies of comprehensive genomic sequencing in healthy newborns, directed by members of her mentorship team. The proposed training will make the candidate a well-rounded ELSI researcher able to deploy mixed methodologies while leveraging her technical background, preparing her to contribute to NHGRI’s goal of developing and assessing strategies for implementing the use of genomic information at the population level. PROJECT NARRATIVE If healthy babies are sequenced near birth, when should genetic information relevant to later stages of their lives be revealed? There are different possible strategies for when this genomic information is revealed, and this research will determine their clinical feasibility, analyze their ethical, legal and social implications, understand diverse parents' preferences, and seek consensus on the necessary and desirable features of a strategy. The project will generate concrete recommendations for implementers of newborn sequencing and will hence help ensure the ethical rollout of genomic medicine for all. | Other Research-Related | 6project_grants_public |
gen_861f35591d10f3255ff6da55620a4d3a | Continuum of Immune Responses to Cryptococcus neoformans | NIH | UNIVERSITY OF MINNESOTA | 1R01AI176922-01 | Abstract Cryptococcal meningitis (CM), caused by the fungal pathogen Cryptococcus neoformans (Cn), is among the most prevalent HIV/AIDS-associated opportunistic infections and causes 15% of AIDS- related mortality globally. In healthy individuals, exposure to Cn in early childhood results in a pulmonary latent infection that is asymptomatic, but leads to the formation of lung granulomas. Following HIV-associated compromise of the immune system, control of latent Cn infection within pulmonary granulomas is lost and the fungus disseminates to cause meningitis. Most studies examining host-pathogen interactions in Cn are observational studies in human cohorts or analyze reference Cn strains in acute disease models of cryptococcosis. We recently showed that the mouse model accurately recapitulates differences in human survival that are observed across Cn clinical isolates and used these data to develop a mouse model of latent Cn infection. Our preliminary data using these mouse models to analyze the immune response to over 50 Cn clinical isolates from individuals with advanced HIV revealed a continuum of disease outcomes that we classified into 3 groups: 1) latent infection resulting in granuloma formation and control; 2) lethal disease similar to that observed with Cn reference strains; and 3) hypervirulence resulting in rapid mortality. Previous studies with reference strains revealed lethal disease is associated with various Cn virulence factors and a detrimental host Th2-mediated type-2 immune response. In contrast, disease prevention is associated with the type-1 cytokine IFNγ. How these Cn-host interactions differ to cause the continuum of disease observed in immunocompromised individuals with HIV is not well defined. We will use analysis of clinical isolates in mouse models of disease to test our central hypothesis that antigenic differences between Cn clinical isolates lead to either protective or detrimental immune responses in the host. We will test this hypothesis by pursuing three specific aims. Our first and second aims will determine the host cellular and effector functions that result in either latency (Aim 1), lethal disease (control infections), or hypervirulence (Aim 2). Our third aim will identify Cn gene alleles for antigens that influence the immune response and ultimately disease outcome. Taken together, these translational studies will define the molecular processes underlying the continuum of Cryptococcus disease with the goal of developing novel immune-modulatory treatment strategies for at-risk patient populations. Project Narrative Cryptococcus neoformans is a fungal pathogen that causes cryptococcal meningitis, a severe HIV-associated infection of the central nervous system. In healthy individuals, C. neoformans establishes latent infections that reactivate and cause clinical symptoms upon HIV-associated immunosuppression, yet disease manifestation differs across patients. Using a mouse model, this research will determine the continuum of patient immune responses and fungal genetic factors that influence clinical outcome towards the goal of developing novel immune-modulatory treatment strategies. | Non-SBIR/STTR | 6project_grants_public |
gen_1ab3e0be631f9ad1c154b64ba033fc7e | A platform to identify in vivo targets of covalent cancer drugs in 3D tissues | NIH | SCRIPPS RESEARCH INSTITUTE, THE | 5R33CA281918-02 | Abstract Covalent inhibitors represent some of the most successful drugs in human history, including aspirin and penicillin. Recently, targeted covalent drugs have taken center stage as a compelling approach for achieving major goals in oncology that have proven elusive for more classical reversible small molecules, including, for instance, the selective inactivation of oncogenic kinases (BTK, EGFR, FGFR, JAK3) and, most notably, the inhibition of the once-deemed undruggable KRAS protein. We are now in the midst of a resurgence of interest in covalent drugs for their demonstrated capability to engage cancer targets that have been historically considered undruggable. However, despite their proven success and inherent advantages of potency, there has been a general reluctance to develop covalent drugs due to the concern of potential irreversible off-target toxicity across different organ systems. Hence, a comprehensive understanding of both on and off-targets in vivo is critical for covalent drugs. Currently, it is impossible to determine drug binding across a whole animal with cellular and molecular resolution in mammals. Building upon a recent breakthrough in tissue imaging termed CATCH (Clearing-Assisted Tissue click Chemistry), we propose to develop a general platform for in vivo imaging of drug-target interactions with unprecedented spatial precision by integrated applications of high-resolution whole-body imaging and chemoproteomics (such as Activity-Based Proteomic Profiling, or ABPP) through the same covalent probes. This way, every cell in a living mammal targeted by the drug (both on- and off-target) can be revealed in situ and registered onto a defined protein map to screen and identify in vivo drug targets. The data stream generated by this platform could rapidly link the rich knowledge of drug affinity to the therapeutic index, therefore accelerating the translation of chemical activities into cancer therapies. Our team has well-established and complementary expertise in chemoproteomics and tissue imaging to ensure the successful execution of the project. In this IMAT R33 application, we plan to further develop CATCH to profile in vivo targets of covalent kinase inhibitors. First, we will adapt CATCH to 3D somatic tissues (Aim 1). Next, we will expand CATCH to an array of covalent BTK (Bruton’s tyrosine kinase) inhibitors (Aim 2). Finally, we will profile dose-dependent in vivo cellular targets of BTK inhibitors in the mouse cardiovascular system (Aim 3). We anticipate that these studies will establish in vivo CATCH methods for identifying targets of covalent BTK inhibitors to better understand their efficacy and toxicity. More generally, the established platform can be broadly applied to any covalent cancer drug for unbiased in vivo target identification. The pipeline, analytics, and high-resolution drug target data will be rapidly disseminated for public access and exploration, releasing an immediate, direct, and profound impact on covalent cancer drug discovery and refinement. Project Narrative Recent breakthroughs in targeted covalent inhibitors are revolutionizing the treatment and prognosis of many cancers, however, people have been reluctant to develop new covalent inhibitors due to the fear of off-target toxicity. This proposal aims to combine chemoproteomic and tissue imaging approaches to establish a 3D, whole-body platform for comprehensive identification of on- and off- targets of covalent drugs in vivo. The resulting technologies and data will have an immediate and transformative impact on the development and refinement of cancer therapies. | Non-SBIR/STTR | 6project_grants_public |
gen_e2eea12f200ef73100e35be627b4bdd4 | 5T-IV: photoacoustic needle with beacon pulse for ultrasound guided vascular access with Tool-Tip Tracking and Tissue Typing | NIH | JOHNS HOPKINS UNIVERSITY | 5R61HL168779-02 | Project Summary Vascular access is the most common medical procedure in the world, with over a billion insertions performed annually. In the United States, over 350-million peripheral intravenous (IV) catheters are sold yearly, in addition to millions of central and arterial lines. However, peripheral IV placement often requires several attempts at insertion, which is painful and time consuming. Furthermore, there is a high rate of early failure that can lead to further complications, including extravasation, thrombophlebitis, and compartment syndrome, as well as delays in delivery of therapeutic medications. Ultrasound guided vascular access is widespread and has advanced difficult IV access success rates, but there is clearly room for improvement as the success rate for ultrasound guided peripheral IV placement is stagnant at 80%. Improving the success rate for vascular access will have a huge impact on patient care and patient safety. To address the clinical need described above and overcome the current technical challenges, we have pioneered a novel concept of ‘active acoustic communication’ between needle and imaging systems for guidance. We are further evolving the concept into a clinical prototype that is intuitive and easy to use, low cost, disposable, works with any ultrasound vendor, offers easy needle tip tracking, and differentiates between tissue types. A ‘5T (tool tip tracking + tissue typing)-IV’ guidance platform will offer an integrated interface layer for clinicians, providing (1) high-spatiotemporal resolution (sub-millimeter and sub-second) tool tip tracking over a wide field-of-view, (2) encoded contrast with a flashing active echo pulse to visualize the needle tip, and (3) highly accurate tissue typing at the needle tip with (4) an unaltered clinical workflow. In this project, we will first develop a vendor-independent clinical prototype hardware and software package with comprehensive performance characterizations in tissue-mimicking phantoms, and this will guide further iteration of the form- factor and software user interface included in the clinical prototype to be tested in animals. The efficacy and user experience will be extensively evaluated in preclinical phantom models by performing human factor studies with 65 end users. Quantifiable metrics (e.g., success rate, time duration and number of trials) will be compared between conventional US and the 5T-IV guidance methods will be expanded to swine animal models for both jugular and femoral cannulation with different experience level (residents vs. attending anesthesiologists). Project narrative Vascular access is the most common medical procedure in the world, with over a billion insertions performed annually. Ultrasound-guided vascular access is increasingly utilized but still has success rate stagnant at 81% primarily due to low contrast resolution, and the unsuccessful cases can develop significant patient discomfort, decreased efficiency, increased costs, and occasionally life and limb threatening complications. To address the enormous problem, we propose a vendor-independent ‘5T (tool tip tracking + tissue typing)-IV’ guidance platform to augment the clinical US imaging guidance. | Non-SBIR/STTR | 6project_grants_public |
gen_e7db4ad5196484dc066c6d072cbf443b | Role of Vitamin C in Low back Pain and Intervertebral Disc Degeneration | NIH | UNIVERSITY OF MINNESOTA | 1R21AT012559-01A1 | PROJECT SUMMARY / ABSTRACT Persistent back pain is a leading cause of disability worldwide and one of the most common reasons pa- tients are prescribed opioids, despite their poor ability to improve function. Chronic back pain threatens our economic health due to high rates of health care utilization and disability and our quality of life due to pain-related suffering, pain-associated opioid misuse, depression, and anxiety. The primary driver in 40% of all low back pain (LBP) cases is estimated to be pain from intervertebral disc (IVD) degeneration. Ex- isting symptomatic (e.g., analgesics) or invasive (e.g., spinal injections, surgery) treatments have limited efficacy and are associated with risk for opioid misuse or surgical complications, respectively. There is an urgent need for safe and effective treatments for chronic LBP that can be rapidly translated to the clinic. Disc degeneration is associated with decreased production of extracellular matrix (ECM) components including collagen and proteoglycans. DNA methylation is an epigenetic mechanism that regulates gene expression and can be modified in response to local, environmental, or pharmacological factors. We have previously shown that a non-invasive treatment (running exercise) attenuates behavioral signs of chronic LBP, increases IVD collagen production, and decreases IVD global methylation. Ascorbic acid (vitamin C) is essential for collagen production and is involved in the regulation of DNA methylation. Ascorbic acid is safe for daily consumption and its deficiency is correlated with musculoskeletal pain in elderly. Although ascorbic acid exhibits analgesic properties in some conditions and is essential for colla- gen synthesis and musculoskeletal health, its therapeutic potential for LBP has not been studied. The overall objective for this application is to determine the therapeutic outcomes of ascorbic acid treat- ment in discogenic LBP and IVD degeneration using a pre-clinical model of progressive IVD degenera- tion associated with LBP. Our central hypothesis is that ascorbic acid reprograms expression of ECM genes by decreasing DNA methylation in degenerated IVDs, thereby attenuating discogenic LBP. In Spe- cific Aim 1, we will examine the therapeutic efficacy of long-term ascorbic acid supplementation (4 months) on the prevention and treatment of LBP and disc degeneration using a mouse model. In Specific Aim 2, we will determine the effects of vitamin C on DNA methylation and mRNA expression of ECM genes, with emphasis on collagens and proteoglycans, and will correlate these data to disc pathology and behavioral signs of LBP. The expected outcomes are that we will observe therapeutic effects of ascorbic acid on LBP and IVD degeneration (Aim 1) associated with reprogramming of IVD DNA methyl- ation (Aim 2). These results will provide critical proof-of-concept data for using noninvasive, safe, dietary vitamin C supplementation as a treatment for discogenic LBP. PROJECT NARRATIVE Persistent back pain is a leading cause of disability worldwide and is frequently caused by damage to the intervertebral discs in the spine. The goal of this project is to generate proof-of-concept data examining the efficacy of vitamin C as a dietary supplement for the treatment of low back pain. In addition, we will test the hypothesis that vitamin C treatment alters extracellular matrix gene expression in degenerated intervertebral discs. | Non-SBIR/STTR | 6project_grants_public |
gen_014b9017eb9b5ef3517577517ad5a57c | Macro-level Health Considerations of Community and Criminal Justice System Relationships in North Texas | NIH | HEALTHY TARRANT COUNTY COLLABORATION | 3OT2OD035659-01S1 | Policing and incarceration are root causes of health inequity, but to date, there have been few efforts to intervene on law enforcement practices and the carceral system as structural factors shaping poor health. Addressing this gap, we propose a Community-Led Structural Intervention to Reduce Police-Related Health Inequities aimed at reducing discretionary arrests and incarceration Tarrant County, Texas. This study seeks to address the impacts of policing and incarceration on Tarrant County’s disproportionately impacted communities. The structural factors of the criminal legal system, related policy, and the role of police actions in exacerbating mental and physical health outcomes will be investigated throughout the course of the funding process. Our study will bring together a coalition of stakeholders affected by policing, including Black, Latine/Latinx, and LGBTQ+ communities, to identify commonalities and particularities of policing among each community. Using the PRECEDE-PROCEED model of intervention planning, we propose a multi-phase process for identifying changeable priorities related to policing. We expect the specific health outcomes that will be assessed include use of force and poor mental health days. Further, this study will bring together a diverse group of community members through a community advisory board, thereby engaging community members that have lived experiences with mass incarceration and policing in order to improve the population health within the community. Upon successful completion of this work, we expect to have developed, implemented, and evaluated a pilot intervention that addresses policing and incarceration as a structural and political determinant of health that has individual, interpersonal, and community consequences. | Other | 6project_grants_public |
gen_a25cf7c69ae4fe2868a72f69bedc869e | Interplay of processed diet, gut microbiota, and interferon-linked mucosal immunity in the onset and prevalence of inflammatory bowel disease | NIH | PENNSYLVANIA STATE UNIVERSITY, THE | 1R01DK133334-01A1 | PROJECT SUMMARY/ABSTRACT Environmental factors including diet and gut microbiota profoundly impact the host resistance and susceptibility towards intestinal inflammation. A high intake of ultra-processed foods enriched with refined starch, sugar, protein, and hydrogenated fat and low in whole grain contents increases the risk of inflammatory bowel disease (IBD). Despite such robust epidemiological data that ultra-processed food is associated with a higher risk of IBD, the underlying mechanisms by which ultra-processed foods escalate IBD susceptibility remains sparse. This proposal aims to elucidate such mechanisms with a goal to devise a dietary-based intervention(s) to reduce IBD onset and occurrence. Interferon gamma (IFNγ)-inducible immunity-related GTPases (IRGs) family M, named IRGM in humans and Irgm1 in mice, is a disease susceptibility risk allele for Crohn’s disease in humans. Irgm1 orchestrates autophagy-mediated immunity against bacteria and impedes NLRP3 inflammasome activation. Our preliminary observation demonstrated that ultra-processed ingredient diet (PID) reduced colonic expression of IFN-γ inducible genes, which protect against invading microbes. PID-fed mice also exhibited increased encroachment of colonic bacteria into the mucus layer and NLRP3 inflammasome activation. Notably, these mice developed severe experimental acute (induced by dextran sulfate sodium, DSS) and chronic (induced by IL-10 receptor neutralization) colitis. Based on our preliminary data, we hypothesize that heightened microbial encroachment due to impaired host resistance against microbes and persistent activation of NLRP3 inflammasome escalates susceptibility to IBD in PID-fed mice. This hypothesis will be tested by pursuing three specific aims: Aim 1 : Assess the role of IFNγ-inducible GTPases in escalating PID-induced predisposition to IBD. Aim 2 : Assess the role of gut microbiota and their metabolites in the regulation of IFNγ-inducible GTPases and PID-induced IBD susceptibility. Aim 3 : Assess the role of NLR inflammasomes and IFNγ-inducible GTPase axis in PID-induced IBD susceptibility. This proposal has potential implications for public health, given that the prevalence of IBD has risen in parallel with the increase in the intake of ultra-processed foods. Completion of the aims of this proposal will identify the mechanism(s) by which a processed diet increases the risk of IBD. A mechanistic understanding of how ultra-processed diet increases susceptibility to IBD will lead the way toward defining a dietary strategy to reduce the incidence of IBD in humans. 1 PROJECT NARRATIVE It is widely accepted that long-term intake of ultra-processed foods has detrimental effects on host intestinal health; however, the underlying mechanisms by which processed ingredient diet escalates susceptibility to intestinal inflammation are ill-defined, thus preventing us from devising a dietary-based intervention(s) to reduce the onset and occurrence of inflammatory bowel disease (IBD). Our preliminary observation demonstrates that the processed ingredient diet (chiefly comprised of refined starch, sugar, and fat but lacking in whole grain contents) increases the propensity to develop IBD by impairing the host mucosal immune responses against invading (opportunistic) microorganisms in the gut. This proposal seeks to further investigate this exciting novel observation, address the potential underlying mechanisms, and identify processed diet-linked microbial metabolites that escalate susceptibility to IBD. | Non-SBIR/STTR | 6project_grants_public |
gen_63d5ab7ecee63c727dc988dbd9bb0ae2 | Tracing spatial organization of germinal centers in rhesus macaques | NIH | GEORGIA INSTITUTE OF TECHNOLOGY | 5R21OD035574-02 | Germinal centers (GCs) are the microstructural sites in secondary lymphoid organs, but GC structures are impaired in AIDS patients due to the loss of supporting CD4+ T cells, leading the deficiencies in immune responses and even inefficacies in antiretroviral therapies in HIV-infected individuals. To solve this challenge, rhesus macaques are well-established nonhuman primate models (NHPs) to study the immunopathogenesis of HIV. However, the spatial coordination of Follicular dendritic cells (FDCs), T cells, B cells, extracellular matrix (ECM), and cytokine regulation of simian immunodeficiency virus (SIV)+ infected macaques and SIV- macaques is still not clearly understood. Thus, there is a critical need to decipher the spatial and temporal control of (1) cytokine, (2) ECM, and (3) T-B cell interactions at the single cell level in GCs for identifying the (1) spontaneous activation and (2) SIV-infection-induced responses in the immune system of HIV/SIV disease. To shed light on immune regulation in lymph node tissues of SIV+ and SIV- macaques and human donors, this project will leverage spatial proteomic and transcriptional profiling to map B cell subsets and their interactions with T cells, ECM, and cytokines in the lymph node tissues. The long-term goal is to generate single cell insights into B cell development in GCs of SIV+ and SIV- macaques in response to spontaneous activation and infections. The goal of this project is to define spatially resolved cellular interactions and cytokine/ECM gradients pixel-by-pixel in fixed macaque and human tissues. The hypothesis is that (1) spatial distributions of cell types, cytokines, and ECM of spontaneous GC activation are uniquely controlled by CD4/FDC and B cells in NHPs compared to humans, and (2) B-cell responses are spatiotemporally regulated by CD4/FDC cells in NHPs with and without SIV infections. The rationale for this hypothesis is based on the evidence that (1) IL- 10+ cells were spatially close to viral SIV-DNA+ lymphoid cells in SIV+ tissues and (2) heterogeneous GC activation and spatial GC organization maps in macaque and human tissues. The central hypothesis will be tested by pursuing two specific Aims. Aim 1 will evaluate the effect of multiplexed cytokine, ECM, and CD4/FDC interactions with B cell subsets in SIV- macaque (n=24 each) and HIV- human tissues (n=24). Aim 2 will evaluate cytokines, ECM, CD4/FDC, and B cell maturation in lymph node tissues of SIV- and SIV+ macaques (n=24). To accomplish these Aims, spatial cell phenotyping and cytokine gene expression profiling will be used to analyze B cell development and statistical comparisons of spatial cell neighboring features in macaque and human tissues. This project builds an interdisciplinary team integrating experts from spatial omics, NHP immunology and pathology, and bioinformatics. The proposed application is innovative because it uses cutting-edge technology to study spatial proteomics and transcriptomics of lymph node tissues of macaques and humans. This research is significant because it defines spatial GC organization to understand why SIV+ infections induce GC defects and how it deviates from human immune responses. | Non-SBIR/STTR | 6project_grants_public |
gen_239a70a8ad568e1a572685ca4d2e5a17 | A Healthy Weight Intervention for Family Stress during the Early Phases of ALL Treatment: NOURISH-ALL | NIH | UNIVERSITY OF KANSAS MEDICAL CENTER | 1K08CA279877-01A1 | PROJECT SUMMARY/ABSTRACT Excessive weight gain leading to obesity is common during the early phases of pediatric acute lymphoblastic leukemia (ALL) treatment, and results in higher risk of relapse, lower event-free survival rates, and more adverse events during treatment. Yet, effective preventive interventions for excessive weight gain during the early phases of ALL treatment are lacking. Aligned with NCI priorities, the long-term objective of this work is to mitigate weight- related disparities in pediatric cancer treatment outcomes. Guided by the ORBIT Model of Behavioral Intervention Development, the goal of this proposal is to adapt, refine, and pilot test a family-based health promotion intervention that aims to curb excessive weight gain among youth with ALL by integrating support for family coping with stress during the early phases of ALL treatment. Given that ALL is most prevalent in young children, families play an essential role in shaping youth’s health behaviors during treatment. In Aim 1a, the PI and her mentorship team will adapt an existing family-based health promotion intervention (NOURISH-T) to meet the specific needs of families of youth in the early phases of ALL treatment (NOURISH-ALL). Adaptations will incorporate family systems and cognitive behavioral intervention components to support healthy family coping with diagnosis and treatment stress. Additional, patient-centered adaptations regarding intervention content and delivery will be informed by semi-structured input from families and youth with ALL (n=10 at minimum) and multidisciplinary clinical experts (n=6 at minimum) until thematic saturation is reached. In Aim 1b, the investigative team will iteratively refine the NOURISH-ALL intervention through sequential testing with families and youth with ALL (n=5 at minimum) and structured participant feedback. Intervention refinement will be ongoing until >80% feasibility and acceptability ratings are achieved or until n=10 families complete the intervention and provide structured feedback. In Aim 2, the adapted and refined NOURISH-ALL intervention will be pilot tested in a single arm trial with 20 newly recruited families of youth in the early phases of ALL treatment. The research team will assess key components of participant engagement to inform the future, fully powered clinical trial, including recruitment rate, retention at treatment completion, and intended intervention dose received. By incorporating tailored strategies for health promotion during the early phases of ALL treatment, the proposed study seeks to shift clinical practice paradigms to prevent weight-related disparities in treatment outcomes. This K08 will provide opportunities for the PI to acquire skills and knowledge in: (1) the early phases of ALL treatment, (2) scientific adaptation and refinement of family-based behavior change interventions, and (3) the conduct of clinical trials focused on behavior change interventions. The research and career development plan, supported by a multidisciplinary team of experts in a rich academic environment, will support the PI’s transition to independence as a cancer control scientist who possesses the skills and expertise needed to adapt evidence-based behavior change interventions to the pediatric cancer treatment context. PROJECT NARRATIVE Excessive weight gain leading to obesity is common during the early phases of pediatric acute lymphoblastic leukemia (ALL) treatment, and results in higher risk of relapse, lower event-free survival rates, and more adverse events during treatment. To address multiple calls from the NCI and other federal agencies for research targeting disparities in treatment outcomes among patients who experience obesity, this K08 research will adapt, refine, and pilot test family-based health promotion intervention that aims to curb excessive weight gain among youth with newly diagnosed ALL. The proposed research and career development plan will yield important patient- centered data regarding components of intervention adaptation, feasibility, acceptability, and participant engagement, enhancing the potential for the investigator to advance behavioral cancer control science and improve clinical outcomes for youth with ALL. | Other Research-Related | 6project_grants_public |
gen_ac87465e493f699b21a363fe22f1a2d0 | Evaluation of Cancer Health Activism Network for Greater Equity (CHANGE) | NIH | RICE UNIVERSITY | 5R25GM150177-02 | The Cancer Health Activism Network for Greater Equity (CHANGE) project addresses inequities in education and health outcomes for African American students in biosciences by enhancing biology teachers' cancer research and cancer health disparities knowledge and developing and implementing transformative, innovative, socially just biology lessons. The lessons will be refined over a series of Saturday workshops (CHANGE Academies) for students in schools that primarily serve Black and African American students in the Houston area. Additionally, locally and nationally, CHANGE will broadly disseminate these lessons through in-person and virtual professional developments (Health Disparities Institute for Teachers) for educators. The specific aims of this collaboration are to (i) increase teacher knowledge of both cancer biology and cancer health disparities and (ii) produce a novel high school biology curriculum on cancer biology and health disparities research grounded in the Transformative Leadership framework, and (iii) assess the impact of the CHANGE curriculum on student outcomes. The team that we have assembled includes the highest caliber of NIH-funded researchers, leaders in STEM teacher Professional Development (PD), experienced curriculum writers, leading bioethicists, and social scientists with expertise in racial disparities and inequities and large, diverse school districts. The PI and the research team have a track record of providing teacher PD that enhances and improves teachers' science content knowledge, self-efficacy, and leadership, improving student outcomes. Rice University is an ideal location for this project because it is strategically located in the large and diverse city of Houston, Texas, and is adjacent to the Texas Medical Center (TMC), the largest medical complex in the world. These collaborations routinely occur between the TMC and Rice University. This proposal is responsive to the goals of the NIH SEPA program as it will focus on Research Experiences, Curriculum or Methods Development, and Outreach to increase Black and African American interest and motivation in biosciences fields. The outcomes of this project, including the curricular materials that will be developed, will advance current knowledge about effective social justice in high school education as we pursue educational and health equity to successfully promote a diverse bioscience workforce. Cancer Health Activism Network for Greater Equity (CHANGE) aims to increase Black and African American K-12 student motivation and interest in biosciences and health careers by developing and disseminating novel high school biology lessons that focus on cancer biology and cancer disparities with a social justice lens. High school biology teachers will participate in research internships to study cancer fundamentals and will work with a team of experts to develop lesson plans that connect biology standards with cancer biology and disparities research. We hypothesize that aligning cancer research with the Transformative Leadership framework will create equitable learning experiences for Black and African American students, motivating them to pursue bioscience careers. | Other Research-Related | 6project_grants_public |
gen_e99f4effd9db13145ed99d5392bb9d93 | Smart Cuff: Multi-Parameter Hemodynamic Monitoring via a Single Convenient Device | NIH | UNIVERSITY OF PITTSBURGH AT PITTSBURGH | 5R01HL163691-02 | PROJECT SUMMARY/ABSTRACT Multi-parameter hemodynamic monitoring is needed to manage surgical and intensive care patients. Monitoring blood pressure (BP), cardiac output (CO), and left ventricular ejection fraction (EF), in particular, permits detection of frequent hypotension and hemodynamic instability, diagnosis of the cause for selecting appropriate therapy, and titration of interventions (e.g., goal-directed therapy). However, measurement of these three hemodynamic variables currently requires multiple devices that are invasive, manual, or specialized. While the oscillometric arm cuff device is non-invasive, automated, and standard, it only estimates BP from the measured cuff pressure waveform via a population average algorithm that does not maintain accuracy over the clinical range. The overall goal of this project is to extend the ubiquitous arm cuff device for accurate and convenient multi- parameter hemodynamic monitoring via smart algorithms. The specific aims are: (1) to build an arm cuff device for recording cuff pressure waveforms; (2) to simultaneously acquire patient data with this and reference devices for algorithm training; (3) to develop and incorporate algorithms for accurately computing BP, CO, and EF from the cuff pressure waveform based on the training data; and (4) to validate the real-time Smart Cuff against reliable reference measurements in patients. The device will be developed to control the cuff pressure and incorporate custom algorithms. The cuff pressure waveform via the device and reference BP, CO, and EF via arterial and pulmonary artery catheters and echocardiography will be recorded before and after clinical interventions in many surgical and intensive care patients. These training data will be analyzed to refine or adapt previous physiologic algorithms and to investigate potentially superior machine learning algorithms for best estimation of the three hemodynamic variables. The final algorithms will be implemented for a real-time device, and the integrated system will be tested against the same reference measurements during clinical interventions but from new patients. Achievement of the specific aims will be followed by a translational project to bring the Smart Cuff to patient care and a research project to extend the device capabilities including addition of automated clinical decision support. Ultimately, these efforts may help in improving patient outcomes and reducing healthcare costs in the near-term. PROJECT NARRATIVE Comprehensive cardiovascular monitoring is needed to care for very sick patients. However, such monitoring currently requires multiple devices that are invasive, manual, or specialized. In this project, the standard automatic arm cuff device will be extended through innovative algorithms to measure blood pressure with superior accuracy, blood flow, and cardiac function, and the new device will then be evaluated for accuracy in patients. | Non-SBIR/STTR | 6project_grants_public |
gen_ee63369ca4023619612b9d691380b440 | Targeting RAGE in tumor and TME to oppose inflammation and drug resistance in obesity associated ER+ breast cancer | NIH | GEORGETOWN UNIVERSITY | 1R01CA276587-01A1 | PROJECT SUMMARY/ABSTRACT Obesity is associated with increased postmenopausal estrogen receptor-positive (ER+) breast cancer (BC) risk and a 2-4 fold increase in mortality from all BC subtypes. Mechanisms of increased resistance to therapy and ensuing fatal BC metastasis in obesity remain unclear. Here, we study how the inflammatory state of obesity drives ER+ BC. We study how postmenopausal estrogen, estrone (E1), which is 3-fold higher in obesity, co- operates with NFκB and the Receptor for Advanced Glycation End-products (RAGE) to upregulate metastasis. Our data indicate that BC cell:adipocyte contact activates NFκB and E1:ERα to induce pro-inflammatory cytokine genes in both cell types, stimulating greater cancer stem cell expansion, and rapid ER+ BC growth and metastasis. Preliminary data show NFκB and E1:ER co-stimulate genes encoding RAGE-ligands (S100A8/A9). RAGE is a major NFκB activator in other cell types (including immune and endothelial cells), but little is known of RAGE/NFκB signaling in BC. We showed that RAGE acts in both tumor cells and the host microenvironment (stroma and fat) to upregulate cytokines that recruit myeloid-derived suppressor cells (MDSC) to promote metastasis. In obese mice, E1 increases BC growth, in part by stimulating immune evasion. New data show antiestrogen-resistant ER+BC lines, including those bearing ESR1 mutations, show increased NFκB activity and RAGE levels. Moreover, the investigational RAGE inhibitor, TTP488, cooperates with the ER-blocker, fulvestrant, to arrest antiestrogen-resistant ER+ BC cell growth in multiple resistant lines. Here, we test if E1-bound ER, NFκB, and RAGE interact in ER+ BC, adipocytes and immune cells to drive gene programs of endocrine therapy resistance and if RAGE inhibitors reverse this. We hypothesize that E1:ER and NFκB cooperate to induce RAGE, and RAGE activates NFκB in ER+ BC tumor cells and peritumoral fat to drive pro-inflammatory, pro-metastatic gene expression programs of tumor progression and acquired drug resistance. Aim 1 will identify tumor cell-intrinsic feed-forward mechanisms mediating RAGE/ NFκB activation in an E1-rich breast cancer environment, testing if E1 and NFκB induce RAGE to activate Rac1, and TLR4 driving feed-forward oncogenic NFκB activation in ER+ BC. Aim 2 will test if RAGE mediates pro-oncogenic, pro-inflammatory target gene activation by E1/ER and NFκB in breast cancer cells. We will identify E1/ER and NFκB cistromes and transcriptomes and test if these require RAGE. The relevance of RAGE-dependent ER/κB co-target genes activation in obesity will be validated by comparing ScRNAseq in human ER+ breast cancers from obese and lean donors. Aim 3 will identify tumor cell-extrinsic mechanisms whereby peritumoral fat in obese hosts promotes acquired antiestrogen resistance and immune evasion in ER+ cancers. Aim 4 will test if RAGE inhibitors restore endocrine therapy responses in organoid and PDX models derived from ER+ breast cancers. This work could identify new approaches to treating acquired endocrine resistance in metastatic ER+BC, particularly in obese patients. Project Narrative Obesity is associated with increased estrogen receptor-positive (ER+) breast cancer (BC) risk and a 2-4 fold increase in mortality across all BC subtypes, but mechanisms of increased acquired resistance to therapy and ensuing fatal BC metastasis in obesity remain unclear. This project studies how the estrogen, estrone (E1), which dominates after menopause and is much higher in obese local mammary fat, co-operates with ER/ NFκB and the Receptor for Advanced Glycation End-products (RAGE) in BC cells, adipocytes, and the tumor microenvironment to impair drug treatment responses. Preclinical studies in organoids and PDX models test the ability of new RAGE inhibitor drugs to reverse acquired endocrine therapy resistance. | Non-SBIR/STTR | 6project_grants_public |
gen_a5d5c02030bd093c46f5900cbcbff289 | Augmenting cancer checkpoint immunotherapies via microbially-derived metabolites | NIH | UNIVERSITY OF CALIFORNIA-IRVINE | 1K22CA272915-01 | PROJECT SUMMARY/ABSTRACT Immune checkpoint inhibitors (ICIs) have significantly improved long-term survival across diverse cancer types including melanoma, non-small cell lung cancer, triple negative breast cancer, and others. However, ICI efficacy relies on multiple cancer, host, and environmental variables, and only a small fraction of patients will respond to these antibody drugs. Methods to improve ICI responsiveness are therefore a highly desirable, unmet clinical need. Human-associated microbes are critical regulators of host health and disease including cancer treatment. Clinical studies have shown that specific gut bacterial species correlate with improved patient outcomes of ICI therapy, and colonization by these active microbes can directly elicit antitumor activity in preclinical animal models. These observations raise a fundamental question: what are the microbial mechanisms that dictate ICI efficacy? My previous work has demonstrated that a secreted bacterial peptidoglycan hydrolase is sufficient to broadly improve ICI therapy in murine models of cancer. Moreover, this phenotype could be recapitulated simply by coadministration of a synthetic fragment that mimics the product of the peptidoglycan hydrolase. These findings raise the exciting hypothesis that the production of microbial metabolites can directly improve ICI efficacy. The main objective of my proposal is to examine enzymatic mobilization of bacterial PG metabolites as a general mechanism of immune modulation during cancer ICI therapy. Aim 1 will explore host enzymes as new factors that determine ICI efficacy. Aim 2 will produce chemical probes to discover ICI-activating bacterial enzymes. Aim 3 will examine PG mobilization as a broad-spectrum strategy to potentiate ICI response in new indications and against new checkpoint targets. To accomplish these goals, I have built a broad and interdisciplinary skill set from my graduate work in chemical tool development with Dr. Linda Hsieh-Wilson at Caltech and my postdoctoral work in host-microbial communication and cancer immunology with Dr. Howard Hang at Scripps Research. To complement these strengths, I have established collaborations with leaders in the fields of cancer immunotherapy and host-microbial interactions to provide training in new cancer model systems and access to critical human-derived isolates, which will greatly aid in my efforts to establish the generality and human relevance of PG mobilization during ICI treatment. In addition, I have proposed a comprehensive career development plan to address any residual gaps in my abilities to effectively manage a laboratory, disseminate our findings, and obtain independent funding. The acquisition of these skills during the K22 period will fuel progress towards the completion of my proposal, providing key preliminary data needed for my first NCI R01 grant application. My scientific and career development enabled by the K22 award will help me to achieve my long-term career goal to become a successful independent investigator at the intersection of host-microbial communication and cancer immunotherapy. Moreover, these efforts may yield mechanistic insights and translational avenues to understand and augment differential ICI responses in the clinic. PROJECT NARRATIVE Cancer immunotherapies including checkpoint inhibitors have revolutionized the treatment of multiple cancer types, yet these drugs are only effective in a small subset of patients. Although specific members of the gut microbiota have been positively correlated with patient outcomes, the mechanisms underlying these clinical observations remain largely unknown. Using a combination of chemical tools, biochemical characterization, and in vivo model systems, this proposal will examine enzymatic mobilization of microbial glycans from the gut as a common mechanism to prime host immune responses during checkpoint blockade, with the goal of augmenting the efficacy of current and next-generation checkpoint immunotherapies. | Other Research-Related | 6project_grants_public |
gen_c1980a1637ccce3868daf537960297c7 | Temporal trends in quality indicators of palliative care for patients with chronic illness hospitalized with acute respiratory failure | NIH | UNIVERSITY OF WASHINGTON | 5F32HL167667-02 | PROJECT SUMMARY/ABSTRACT Patients with chronic illness who develop acute respiratory failure face significant morbidity and mortality, with survivors often suffering serious physical and psychological sequelae. These patients have a critical need for palliative care. Palliative care is focused on improving quality of life for both patients and family of patients with serious illness by providing high-quality communication, symptom control, and emotional and spiritual support. Despite how important this type of care is, there is little information available regarding the quality of palliative care provided to chronically ill patients with acute respiratory failure. Without this knowledge, it is extremely challenging to develop effective interventions to enhance patient- and family-centered outcomes for this population. To fill this key knowledge gap, I will use explicitly defined, measurable items that detail the processes, outcomes, and structure of palliative care to achieve my primary objective of examining temporal trends in palliative care over the last decade, including an assessment of changes that have occurred since the inception of the COVID-19 pandemic. I have identified a cohort of over 16,000 patients with chronic, life-limiting illness hospitalized with acute respiratory failure between 2012 and 2022. Using this cohort, I will achieve my primary objective through two specific aims. Aim 1 will examine temporal trends in quality indicators of palliative care for patients with chronic illness hospitalized with acute respiratory failure between 2012-2022. Aim 2 will assess differences in quality indicators of palliative care for patients with chronic illness hospitalized with acute respiratory failure, comparing patients with COVID-19 to those without COVID-19, between 2020- 2022. By accomplishing the proposed aims, I will identify improvement, stagnation, and decline in care over time in specific process and outcome metrics across multiple palliative care domains, highlighting significant changes occurring during the COVID-19 pandemic and identifying differences in quality indicators of palliative care for those with and without COVID-19. Results from the proposed project will inform my future research to describe mechanisms underlying these trends, with the goal of developing effective interventions to improve care delivery for adults with chronic, life-limiting illness and acute respiratory failure. PROJECT NARRATIVE Patients with chronic, life-limiting illness and acute respiratory failure are at high risk for poor patient- and family-centered outcomes and represent a patient population in need of high-quality palliative care. This project will address our limited knowledge regarding palliative care for these patients and their family members by examining temporal trends in quality indicators of palliative care. Knowledge gained from this study will guide future research to improve palliative care delivery and outcomes for patients with acute respiratory failure and their family members. | Training, Individual | 6project_grants_public |
gen_4952cf517db0051d57faaac4ed8b2734 | Addressing perinatal mood and anxiety disorders (PMADs) through a doula intervention | NIH | UNIVERSITY OF MONTANA | 5P20GM130418-04 | Perinatal mood and anxiety disorders (PMADs) have intergenerational negative impacts on the physical and mental health of women and their families and are the most frequently experienced childbirth complication. PMADs are particularly common in rural populations, who experience additional obstacles in accessing maternal and child health (MCH) and mental health services. Doulas, non-medically trained childbirth and postpartum companions, have been proposed as one way to address existing healthcare gaps, and as a way to improve mental health outcomes, along with additional MCH outcomes such as decreased substance use, increased prenatal and postpartum healthcare visit attendance, and improvements in overall maternal physical wellbeing. The aims of this research are: 1) Explore the use of doulas to improve self-efficacy and maternal mental health outcomes for rural populations; 2) Develop a doula intervention to improve self-efficacy and maternal mental health in rural populations; and 3) Pilot a doula-led intervention to improve self-efficacy and maternal mental health outcomes. Using the NIM....'s experimental therapeutics approach, I will explore the underlying mechanism impacting these outcomes by focusing on how doulas improve self-efficacy among their clients. Following Whitbeck's (2006) model for adaptation of interventions, and community-based participatory research (CBPR) best practices, I will use formative and exploratory research approaches to address knowledge gaps about the use of doulas to improve PMADs and secondary maternal and infant health outcomes. Through this project we will first collect primary qualitative data regarding the feasibility of using doulas to improve PMAD health outcomes and will identify the needs and barriers in accessing mental health care, as well as social and emotional support that could be addressed by doulas (N=40). The proposed research will then synthesize quantitative data on the use of doulas and peer-support models with qualitative data collected from semi-structured interviews to develop and test a doula-led intervention through adapting an existing doula-training. A mixed-methods approach will be used to analyze findings from study participants (N=75) and doulas and providers (N=20). This project will provide preliminary data for the cultural adaptation of the intervention for use with other marginalized groups and rural populations, including Indigenous communities, who experience heightened PMADs and other negative MCH outcomes. In addition to being extremely common, PMADs are frequently undiagnosed and are negatively associated with a broad range of negative outcomes. Of particular concern are findings that PMADs are increasing. Rural populations are estimated to have PMADs rates that are more than double those in non-rural areas and higher suicide rates.These negative MCH health outcomes are particularly prominent in Montana. Research indicates that the use of doulas can improve a wide-range of pregnancy and | Research Centers | 6project_grants_public |
gen_546bf726bb35d6a8385ba6b75967aea6 | Geospatial food access as a driver of environmental oxidant stressors and early obesity | NIH | NEW YORK UNIVERSITY SCHOOL OF MEDICINE | 1K23ES035461-01 | PROJECT SUMMARY/ABSTRACT Racial/ethnic and socioeconomic disparities in obesity begin early in life. Excessive caloric imbalance in pregnancy is a leading suspected contributor to early weight disparities. Yet, prior studies have not accounted for how limited food access may also drive chemical and psychosocial exposures that influence developmental cardiovascular and metabolic programming to increase early childhood obesity risk. This Mentored Patient- Oriented Research Career Development K23 proposal investigates how geospatial food access and environmental oxidant stressors contribute to childhood obesity, a serious public health challenge and priority of the National Institute of Environmental Health Sciences. Simultaneously, it will prepare the candidate, Carol Duh-Leong, MD, MPP, to become an independent investigator who will address neighborhood disparities that influence the social and chemical context of early child obesity. This study leverages longitudinal data from the NYU Children’s Health and Environment Study, a participating birth cohort (50% Hispanic, 8% Asian, 7% Black; 57% Public Insurance) in the NIH ECHO program that follows pregnant people prenatally and their children postnatally through early childhood. The candidate proposes to geocode participant home addresses and food environment data to longitudinally model joint relationships among geospatial food access, environmental oxidant stressors, and early childhood obesity outcomes. Specific aims are: 1) Examine whether limited geospatial food access increases phthalate and bisphenol exposures in pregnancy; 2) Examine relationships between limited geospatial food access, psychosocial stressors, and oxidative stress in pregnancy; and 3) Evaluate joint effects of geospatial food access and environmental oxidant stressors across pregnancy and infancy on early childhood obesity outcomes. Closing this gap in knowledge would inform built environment investments and neighborhood strategies to interrupt environmental oxidant stressors and decrease obesity risk through the life course. Through the execution of these aims, the candidate will pursue the following mentored training goals: 1) Advanced geospatial analysis; 2) Environmental exposure and biomarker assessment; 3) Longitudinal analysis and environmental mixture modeling. This proposal draws upon world-class clinical, research, and teaching resources available at NYU Grossman School of Medicine for the candidate’s research and training aims. The candidate has assembled an expert mentorship team of content experts in environmental epidemiology, geospatial analysis, exposure and biomarker assessment, and advanced statistical methods to guide her research and training. This team will provide on-site training tailored to the proposed research aims and will guide the candidate’s transition to independence and her establishment of a research program that applies an interdisciplinary life course approach towards understanding the social and environmental context of early child obesity. PROJECT NARRATIVE The consequences of early childhood obesity disparities are lifelong, with substantial cardiovascular risk to racial/ethnic and socioeconomic minority groups. This study integrates longitudinal geospatial, chemical, and psychosocial data to investigate how limited geospatial food access in pregnancy may increase oxidative stress and childhood obesity risk through mechanisms that involve chemical and psychosocial stressors. Understanding the currently unknown role of geospatial food access in driving oxidant stressors starting in utero would inform built environment investments and neighborhood strategies to decrease obesity disparities across the life course. | Other Research-Related | 6project_grants_public |
gen_4240916a3f15a2703b3505801fdc816a | Interplay between reverse transcription and host restriction | NIH | UNIVERSITY OF ILLINOIS AT CHICAGO | 5R01AI174538-02 | Infectious agents have infected prokaryotes and eukaryotes throughout evolution. Indeed, there is co-evolution of organisms and their infectious agents, with development of protective responses in the hosts and adaptive countermeasures by the infectious agents. Infectious endemic retroviruses like murine leukemia virus (MLV) have existed in mice for millions of years and provide us with an outstanding model system to understand how mammalian hosts suppress virus replication and conversely, how viruses counteract this restriction. One system of viral restriction is conferred by the apolipoprotein B mRNA editing enzyme, catalytic peptide 3 (A3) family of proteins, which are packaged into retroviruses in virion-producing cells and after infection of target cells, either block reverse transcription or deaminate deoxycytidine residues in single-stranded DNA, resulting in uracils and G-to-A mutations in the viral genome. The products of retrovirus reverse transcription (ssRNA, ssDNA and dsDNA) are also sensed by host nucleic acid sensors. Sensor binding to viral nucleic acid leads to the production of anti-viral cytokines and chemokines, such as type I interferons, which “warns” surrounding cells to arm themselves against infection by producing proteins such as A3. These host anti-viral events are believed to occur largely in the cytoplasm, where A3 proteins and many host sensors are believed to function. Retroviruses enter cells when the viral and host membranes fuse and capsids are deposited in the cytoplasm. Reverse transcription initiates from within the capsid and capsid dissociation and reverse transcription are mutually dependent; because DNA is more rigid than RNA, without capsid dissociation, reverse transcription cannot proceed and conversely, the generation of DNA facilitates capsid dissociation. The reverse transcription complex not only consists of viral RNA, DNA and the viral proteins reverse transcriptase and integrase, but viral capsid and other proteins such as the MLV protein p12, which is needed for tethering of the proviral DNA to host chromatin to achieve integration. Recently, there has been much debate as to whether reverse transcription occurs solely in the cytoplasm or in the nucleus or both. Our lab pioneered the use of in vivo mouse models to study how A3 proteins restrict retrovirus infection and has used A3 and nucleic acid sensor knockout (KO) mice and genetically engineered animals that express human A3 proteins in these studies. Our data, based on our analysis of A3 KO mice and cells, suggest that the initial step occurs in the cytoplasm but that reverse transcription may also occur in the cytoplasm. With these mouse models, we have the tools to carry out in vitro, ex vivo and in vivo studies to determine how A3-mediated restriction and sensing of reverse transcripts are integrated with reverse transcription and nuclear entry for MLV and its natural host, the mouse. To accomplish this, we propose 3 aims, that will determine: I. Where in the cells APOBEC3 proteins block reverse transcription or deaminate viral DNA; II. What stage in reverse transcription and where in the cell the host base excision repair enzyme, UNG, removes uracil from APOBEC3G-deaminated viral DNA and the consequences of this for viral escape; III. Whether host sensing of viral nucleic acid takes place in the cytoplasm, nucleus or both compartments. Determination of when and where these events occur is critical to understanding how retroviruses, including HIV, evade host immunity and the identification of which steps are likely to be the best targets for interventional therapies at the early stages of infection. Retroviruses are major causes of disease in animals and humans that replicate by reverse transcribing viral RNA into DNA. The initial immune response to retroviral infection is critical to the ability to clear infection, because once the viral DNA integrates into the host chromosomes, persistent infections arise, leading to diseases such as cancer and immunodeficiencies. Here we will examine the timing and location of virus replication and host response, using unique cell culture and mouse models developed by our lab. | Non-SBIR/STTR | 6project_grants_public |
gen_44a8e084edf8d564e07aff8d44df5a14 | Establishment of a Cell-Based Screening Platform for DNA Encoded Libraries | NIH | UNIVERSITY OF FLORIDA | 1R21GM149993-01 | Project Summary DNA-encoded library (DEL) technology has revolutionized the discovery of protein- binding small molecules. However, traditional DELs, where every molecule is connected to an encoding DNA and the library is produced as an intractable mixture, have not been useful for functional and phenotypic screening. We have been involved in the development of one bead one compound (OBOC) DELs, which can be used for functional and cell-based screens since beads are separable and only a small percentage of the molecules on the beads are linked directly to a DNA encoding tag. To date, the very small number of papers that have described activity-based screens of OBOC DELs have required the use of highly sophisticated microfluidics technology. This project will develop a cell-based screening platform for OBOC DELs that will not require any specialized equipment. If successful, this will make this powerful approach to the discovery of bioactive molecules far more accessible to the chemical biology community. Project Narrative This project will attempt to establish a platform in which one bead one compound (OBOC) DNA-encoded libraries (DELs) can be screened cheaply and efficiently for compounds that affect a biological process of interest in a living eukaryotic cell. If successful, this project will make functional screening of DELs far more accessible to the chemical biology community since it will require no specialized equipment. This should spur the discover of a host of new, bioactive molecules. | Non-SBIR/STTR | 6project_grants_public |
gen_9728fdd121dbbf2cac509bc58725f570 | Integrating liquid biopsy-based epigenetic and imaging modalities to evaluate disease response in multiple myeloma | NIH | UNIVERSITY OF CHICAGO | 1R01CA280637-01 | PROJECT SUMMARY Multiple myeloma (MM) affects ~35,000 adult patients in the United States and causes ~12,000 deaths each year. Novel immunomodulatory drugs and effective multidrug combinations have improved the prognosis for patients, but the vast majority of patients eventually relapse. Even among patients achieve a complete response, ~25% progress within 2 years. Most MM relapses can be attributed to the persistence of measurable (minimal) residual disease (MRD). MRD status has emerged as one of the most important prognostic markers, has therapeutic implications, and is incorporated in the International Myeloma Working Group response criteria for therapeutic response assessment. The gold standard of MRD assessment includes multiparameter flow cytometry and next-generation sequencing (NGS) based on bone marrow aspirates. However, evaluation of MRD using only bone marrow aspirates is prone to false-negatives due to patchy disease involvement, hemodilution of bone aspirate, and extramedullary disease. In addition, bone marrow biopsy is an invasive procedure, hence cannot be performed frequently to monitor MRD. Positron emission tomography/computed tomography (PET/CT) is complementary to bone marrow assessment although gaps remain. We have shown that the 5-hydroxymethylcytosine (5hmC), a stable epigenetic marks generated from active DNA demethylation, in plasma cell-free DNA (cfDNA) could be complementary to PET/CT and was associated with overall survival of patients with MM. Here we propose to apply the highly sensitive mapping of genome-wide 5hmC in cfDNA to identify the optimal combination of serial cfDNA-based 5hmC markers with PET/CT for detecting emergence of MRD. Our central hypothesis is that altered 5hmC signatures in cfDNA are associated with clinically detectable disease by PET/CT and/or NGS, thus offering opportunities for accurate yet less invasive approaches to complement bone marrow-based MRD assessment by NGS. Specifically, we propose to identify 5hmC in cfDNA associated with MRD-positivity (Aim 1), determine cfDNA-based temporal dynamics of 5hmC and fluorodeoxyglucose (FDG)-PET/CT changes associated with MRD (Aim 2), and evaluate performance of different MRD modalities in real-world patients (Aim 3). To address these aims, we have assembled an interdisciplinary team with extensive and complementary expertise. The study leverages the established resources of the UChicago Myeloma Epidemiology Study and our leadership in three clinical trials with banked serial blood samples, bone marrow-paired peripheral blood, and known MRD status by NGS. The knowledge gained from this application may improve clinical utilization of MRD in clinical decision making by proper combination of serial MRD assessments (i.e., cfDNA, PET/CT, and bone marrow) for sensitive tracking of MRD as well as provide novel insights into the epigenetic contribution to MM progression. Identifying patients at high risk of progression after successful treatment will allow for timely implementation of additional therapeutic strategies that may ultimately reduce morbidity and mortality among MM patients. PROJECT NARRATIVE Minimal residual disease (MRD) after upfront treatment of multiple myeloma (MM) is used to identify complete response patients at high risk for progression/relapse, however the standard-of-care for MRD assessment by bone marrow aspirates and imaging are not complete and prone to false-negatives. This work seeks to integrate serial epigenetic profiles in circulating cell-free DNA with imaging data to develop, validate, and clinically translate the composite model for MRD detection and tacking of MRD. Successful completion of this study will allow for timely implementation of additional therapeutic strategies and reduce morbidity and mortality among MM patients. | Non-SBIR/STTR | 6project_grants_public |
gen_e0e7e9c1a4777510ca656ef235c7185a | Mobile Health and Oral Testing to Optimize Tuberculosis Contact Tracing in Colombia | NIH | YALE UNIVERSITY | 5R21AI174129-02 | PROJECT SUMMARY Nearly one-third of the 10 million annual tuberculosis (TB) cases worldwide are missed each year. TB contact tracing is an evidence-based practice that is internationally recommended for finding, treating, and preventing TB among close contacts of TB patients. Contact tracing is also considered a highly significant strategy for advancing the WHO and PAHO goal of eliminating TB in moderate-incidence countries like Colombia by 2050. Unfortunately, large implementation gaps persist. Specifically there is need for 1) better TB screening and testing tools to overcome the low sensitivity of symptom screening and the difficulty of collecting sputum in those without cough, and 2) more client-centered implementation strategies to improve case-finding and linkage to care. In prior work with the Secretariat of Public Health (SPH) in Cali, Colombia, we introduced client-friendly education materials, home sputum collection and transport, and molecular testing for TB. The COVID-19 pandemic has led to further evolution of this service to accommodate telehealth visits, and there is need to further adapt these tools to client expectations and the new digital health landscape. Two of the most promising innovations for achieving these goals are pairing next-generation molecular testing with non-sputum samples like saliva and oral swabs to simplify and improve screening and testing algorithms; and incorporating chat apps (e.g., WhatsApp) and automated customer support tools (i.e., chatbots) as mobile health (mHealth) tools to engage, support, and retain household members. Our scientific approach will be to carry out two cross-sectional studies with nested mixed-methods and design research to refine and adapt these tools for contact tracing. Our overall objectives are to enhance the sensitivity, feasibility, acceptability, and appropriateness of contact tracing, and to prepare for a future household-randomized implementation trial. We propose two specific aims: 1) To determine the diagnostic performance of oral samples for TB molecular testing in possible TB patients in clinic and household settings; and 2) To design and iteratively refine an mHealth strategy for implementing contact tracing optimized for feasibility, acceptability, usability, and appropriateness. The investigative team has expertise in TB diagnostics and clinical care, contact tracing, user-centered design, qualitative methods, and implementation science, and a well-established partnership with the SPH that will ensure an outstanding environment for the proposed research. These studies will determine if oral samples are viable for molecular diagnosis of TB and refine an mHealth strategy for household TB contact tracing, using an innovative design approach that is both theory-informed and community-engaged. This study will ultimately deliver client-centered implementation components for an mHealth contact tracing strategy for future evaluation in a randomized, controlled implementation trial. PROJECT NARRATIVE Contact tracing is a proven intervention for finding undiagnosed cases of tuberculosis (TB) in households of newly diagnosed TB patients, and the proposed study in Cali, Colombia, will evaluate two exciting new strategies to enhance contact tracing. We will first test if TB molecular testing on oral samples is feasible, acceptable, appropriate, and accurate for TB diagnosis in adults and children. We will then determine the feasibility, acceptability, usability, and appropriateness of mobile phone chat apps, including video calls and automated assistants, to arrange and conduct TB contact tracing procedures. | Non-SBIR/STTR | 6project_grants_public |
gen_760a987ba34bdb9a8d938e27ceee0637 | SCH: Simulation Optimization of Cardiac Surgical Planning | NIH | UNIVERSITY OF TENNESSEE KNOXVILLE | 1R01HL172292-01 | Many patients take surgical interventions to fight the battle against heart disease. Surgical successes are critical to the patients’ health and their family well-being. For e.g., atrial fibrillation (AF) is the most common arrhythmia in elder population. Catheter ablation is an established treatment for AF, which sequentially creates incision lines to block faulty electrical pathways. However, there are large variations in surgical outcomes. Modern healthcare systems are investing heavily in sensing and computing technology to increase information visibility and cope with disease complexity. Massive data are readily available in the surgical environment. Realizing the full data potential for optimal decision support depends on the advancement of information processing and computational modeling methodologies. Our long-term goal is to advance the frontier of precision cardiology by developing new sensor-based modeling and simulation optimization methodologies. The objective of this project is to optimize AF ablation by integrating simulation-enabled planning with physics-augmented machine learning of sensor signals from patients who underwent AF ablation. This objective will be accomplished by pursuing 3 specific aims: 1) Physics-augmented artificial intelligence (AI) for cardiac modeling – This approach will assimilate heterogeneous sensing data and incorporate electrophysiology prior knowledge into deep learning to increase the robustness of decision making under uncertainty, thereby driving computer simulation into clinical applications; 2) Optimal sensing and sequential learning of space-time AF dynamics – This approach will provide quantitative knowledge of disease mechanisms instead of subjective knowledge that is difficult to translate (or transfer), thereby reducing healthcare disparity due to the availability of human experts in rural areas; 3) Integrating sensor-based learning and simulation optimization for surgical planning - This approach will integrate physics-augmented modeling (Aim 1) and sensor-based learning (Aim 2) with simulation optimization to improve the clinical practice towards data-driven & simulation-guided surgical planning. This project will make a major breakthrough towards precision cardiology by (i) going beyond the current practice of largely expert-based or ad hoc decisions, (ii) capturing underlying complexities in space-time cardiac dynamics, and (iii) integrating physics-based modeling, sensor-based learning, and simulation-based planning for surgical decision support. This project is relevant to the mission of the National Heart, Lung, and Blood Institute because it will drive cardiac modeling and simulation into clinical applications, promote data-driven and simulation-guided surgical planning, and develop an interdisciplinary workforce who possesses both computational skills and medical insights. The success of this project will make a lasting impact on our society and bring a heart-healthy future. | Non-SBIR/STTR | 6project_grants_public |
gen_b2d39414742fae45b5f1563462fc3da1 | Proteomic and integrative omic profiles of sugar- and artificially sweetened beverage consumption and changes in type 2 diabetes risk factors | NIH | BRIGHAM AND WOMEN'S HOSPITAL | 1K01DK136968-01 | PROJECT SUMMARY: Type 2 diabetes (T2D) is caused both by genetic and environmental factors, such as diet, as well as the complex interactions between them. While diet is the cornerstone for T2D prevention, dietary interventions are often difficult to implement and monitor due to limitations in dietary assessment techniques and strategies to produce dietary changes. Despite efforts to reduce SSB consumption, SSBs remain the largest single source of added sugar in the US. SSB consumption has been linked to a higher risk of T2D and related risk factors, but the underlying biological mechanisms are not completely understood. Proteomic profiling and multi-omic integration allow for more detailed phenotyping that may provide a broader view of diet-associated metabolic changes and their functional interpretation. Examination of plasma proteomic and integrative omic profiles that reflect SSB intake and a common alternative beverage, artificially sweetened beverages (ASB), may enhance current dietary assessment methods and unveil novel biological pathways linking diet to T2D and related risk factors through identification of novel dietary biomarkers. Discovery of plasma proteomic and multi-omic profiles of SSB and ASB consumption has immense potential to provide an objective assessment of individual beverage intake and enable informed beverage choices, which is in line with the precision nutrition approaches emphasized in the National Institute of Health’s (NIH) 10-year strategic plan. This proposal cost-effectively leverages existing proteomics profiling among the Nurses’ Health Study II and Health Professionals Follow-up Study cohorts (n=648). It also examines repeated assays in the ongoing NIH-funded SUBstituting with Preferred OPtions trial, a randomized parallel-arm 6- month beverage trial testing the effects of substituting SSBs with ASB or water among daily SSB consumers. We will utilize proteomic and multi-omic network and machine learning analyses to identify discriminatory profiles between SSB and ASB consumption levels and evaluate the associations of these profiles with T2D risk factors. The central hypothesis is that distinct proteomic and omic profiles reflect habitual SSB or ASB intake and that changes in their omic biomarkers are associated with changes in T2D risk factors, revealing novel biomarkers of beverage consumption and biological pathways modified by beverage consumption. This K01 career development award expands on the applicant’s experience in nutritional epidemiology, omics, and biostatistics to gain proficiency in the design and management of intervention studies, implementation of cutting-edge multi-omic statistical analysis techniques, and scientific leadership for precision nutrition applications for T2D prevention. With mentorship from a renowned multidisciplinary research team, the applicant will gain the crucial skills necessary to advance T2D prevention and refine a framework for the utilization of innovative multi-omics techniques in complementary interventional and epidemiological study designs to inform precision nutrition initiatives and transition to an independent investigator. PROJECT NARRATIVE: The majority of type 2 diabetes (T2D) cases could be prevented or reversed through successful implementation of diet and lifestyle treatment and prevention strategies, yet diet and lifestyle interventions are difficult to adhere to due to an abundance of calories in the environment, which includes calories from sugar sweetened beverages. This proposal presents a unique sub-study within an ongoing randomized controlled trial and existing population-based cohort studies aiming to explore the underlying mechanisms of how sugar- and artificially sweetened beverage consumption influence T2D risk factors; we aim to identify novel biomarkers of dietary consumption that could lead to an individualized, objective assessment of beverage intake to increase efficacy of dietary interventions for T2D prevention. The unique career development plan provides training in the design and management of RCTs, the implementation of cutting-edge multi-omic statistical analysis techniques, and scientific leadership in T2D prevention that complements the applicants current training in nutritional epidemiology, omics, and biostatistics. | Other Research-Related | 6project_grants_public |
gen_6ab100d76c2cd7b2bdfbe0898cf9e631 | An Alternative Pathobiology underlying Severe Asthma | NIH | UNIVERSITY OF PITTSBURGH AT PITTSBURGH | 1K08HL164887-01A1 | Project Abstract This application is for a Mentored Patient-Oriented Research Career Development Award entitled “An Alternative Pathobiology underlying Severe Asthma.” I am an Assistant Professor of Medicine at the University of Pittsburgh seeking additional training in benchtop murine research techniques and bioinformatics and ‘omics analysis to transition from benchtop to translational asthma research. The focus of my research is on the role of Type-1 (T1) inflammation in asthma and how this pathway may contribute to disease severity. While great strides have been made in understanding and treating traditionally described Type-2 (T2) inflammation in asthma with novel biologic therapies, nearly 50% of asthma patients lack evidence of this pathway, and even some with T2 inflammation fail to respond well to T2 therapies. A better understanding of other inflammatory pathways in asthma and how they contribute to disease is essential to identifying novel therapeutics for these patients. My preliminary data have demonstrated the presence of T1 inflammation marked by increased expression of IFN-γ, the chemokines CXCL10 and CCL5 as well as T1 expressing tissue resident memory T-cells (T1 TRM) in ~30% of asthma patients. These patients tend to have more severe asthma, greater exacerbations and higher use of systemic corticosteroids. However, the reliance on bronchoscopy for samples has limited our ability to study the clinical effects of this pathway over time. Furthermore, our understanding of the effects of T1 inflammation on the airways in asthma and the role of T1 TRM cells in maintaining/driving this phenotype are unknown. This study aims to improve our understanding of asthmatic T1 inflammation in three ways: (1) Utilize our T1 dominant murine severe asthma model to assess TRM establishment and reactivation in T1High asthma. (2) To assess the importance of maraviroc (a CCR5 inhibitor) in effecting T1 specific changes on the airway epithelium and mast cell prevalence. (3) The multicenter Severe Asthma Research Program provides clinical data in a large number of asthma subjects with paired sputum samples that will allow us to measure T1 inflammatory markers, assess their correlation with clinical outcomes and identify future biomarkers to better identify T1High asthmatics for future trials. This project will provide unique insight into a novel and poorly understood pathway in asthma with the potential to yield exciting new treatment options for a highly prevalent and severe disease. This study will also provide the opportunity for me to acquire skills in bioinformatics and large ‘omics dataset analysis that will foster my development as a translational physician scientist in severe asthma in addition to advancing my murine model skillset. The work will be carried out at the University of Pittsburgh in the division of Pulmonary, Allergy and Critical Care which has a strong track record of developing physician scientists and boasts a highly developed infrastructure for translational research. I have also assembled a highly accomplished multi-disciplinary mentorship team that is internationally recognized in asthma research, bioinformatics and machine learning. Project Narrative: Asthma is a highly prevalent disease composed of different clinical and biologic phenotypes with variable responses to therapy. We have previously described the presence of Type 1 inflammation in approximately 30-40% of asthma patients which associates with more severe disease and poor response to therapy. This project seeks to expand our understanding of the of the pathologic mechanisms underlying this phenotype as well as the clinical manifestations over time. | Other Research-Related | 6project_grants_public |
gen_025cd18750a27c3d0b05db64203ac8c2 | Structural-Transcriptional Relationships that Improve Y537S Estrogen Receptor Antagonism | NIH | LOYOLA UNIVERSITY CHICAGO | 1R37CA279341-01 | Summary This proposal studies how drug-induced structural changes to Y537S estrogen receptor alpha (ERα) impact anti- tumoral activities in hormone-resistant breast cancer cells. Breast cancer is the second leading cause of cancer death in the United States. Acquired resistance to hormone therapies is a leading contributor to mortality. In approximately 40% of progressive ER+ patients, prolonged selective pressure by antiestrogenic therapies gives raise to tumors bearing activating somatic ESR1 (the gene for ERα) mutations. These mutations resist inhibition by clinically approved hormone therapies and engage new transcriptional programs that boost metastatic potential. Y537S missense mutation is among the most common and enables the greatest hormone-free transcriptional activities and resistance to antiestrogen. Next generation selective estrogen receptor degraders (SERDs) have been clinically deployed to address this mechanism of drug resistance. However, they show variable activities in Y537S ESR1 breast cancers and possess common side-effects that will limit their long-term use. We recently studied how a panel of 17 selective estrogen receptor modulators (SERMs) and SERDs bind to and affect Y537S ERα activities in breast cancer cells. We identified structurally distinct SERMs and SERDs with improved activities in this setting. While structurally distinct, our x-ray co-crystal structures showed that the most effective molecules engaged the same S537-E380 hydrogen bond to reinforce the therapeutic antagonist conformation. Therefore, we hypothesize that novel ligand-dependent structural interactions will improve therapeutic antagonistic activities in the Y537S ESR1 setting. In this study, we will leverage our library of over 100 diverse SERMs and SERDs to reveal the structural-transcriptional relationships that underlie improved anti- cancer activities Y537S ESR1 breast cancer cells. We will start by studying how our library binds to and affects Y537S ERα structure and anti-cancer activities (Aim 1). This approach will reveal the ligand binding modes and structural interactions that enable potency. Next, we will study how the most effective molecules impact Y537S ERα genomic activities including protein-protein interactions, genome binding, and transcriptional programing (Aim 2). This approach will show whether the efficacies of SERMs and SERDs arise from alterations to Y537S ERα genomic activities. Finally, we will reveal the anti-tumor and tissue-specific activities of the most effective SERMs and SERDs in hormone-resistant ER+ breast cancer in vivo (Aim 3). This approach will reveal whether our in vitro observations correspond to improved anti-cancer activities in patient-relevant tumor models. Overall, these studies will provide detailed structural-transcriptional relationships to improve therapeutic targeting of Y537S ERα in hormone-resistant breast cancer. Project Narrative Activating somatic mutations to estrogen receptor alpha (ERα) enable hormone therapy resistance and significantly contribute to breast cancer mortality. We recently identified a novel therapeutic structural interaction that correlates with improved anti-cancer activities for Y537S ERα, the most aggressive and drug-resistant of the known mutants. We will use a library of over 100 structurally diverse ERα-targeted small molecule antagonists to reveal the interplay between Y537S ERα structure, genomic actions, and anti-cancer activities to overcome hormone therapy resistance in breast cancer. | Non-SBIR/STTR | 6project_grants_public |
gen_0bd1b84a2ce8980c6bcdc479cb59a693 | Examining the multilevel factors on quality of end-of-life care among cancer patients in Puerto Rico | NIH | UNIVERSITY OF PUERTO RICO MED SCIENCES | 5P20GM148324-02 | PROJECT SUMMARY/ABSTRACT In the United States (US) and Puerto Rico (PR), cancer is the second leading cause of death. Considering that the population is aging, an increase in the burden of cancer is expected in future years. Consequently, quality of care at the end of life (EoL) is critical for dying cancer patients and their families. Studies in the US have shown that EoL care is expensive for cancer patients and frequently has very low or no clinical value, and often is inconsistent with the patient's needs, values, and preferences. Conversely, timely receipt of palliative and hospice care can improve quality of life and decrease emergency department visits, hospitalizations, and the use of life-extending procedures. Even when progress is made towards improving the quality of EoL care, a growing body of evidence shows disparities among groups within the population. Understanding the different characteristics that affect the quality of care at the EoL is fundamental to implementing effective interventions to reduce inequity and improve the efficiency of the healthcare delivery system. There are few studies in the US and no study in PR evaluating the quality of EoL care from the multilevel perspective. This proposal would be the first of such a series of investigations. PR has a predominantly Hispanic, low-income, and aging population with high insurance coverage. Forty-eight percent of the PR population lives below the poverty level, and there is a shortage of medical specialists, including palliative medicine. This study provides an opportunity to investigate the variations in the use of recommended care, its cost, and the provider and patient's behaviors towards EoL decisions. This study seeks to advance our knowledge and understanding of EoL care among cancer patients by utilizing state-of- the-art claims data analytics coupled with surveys data collection. Towards this effort, we will merge the Puerto Rico Central Cancer Registry-Health Insurance Linkage Database with CMS hospice claims data and conduct physicians, patients, and caregivers’ surveys. Specific Aims are: 1) Assess the use of recommended care and the cost of care among cancer patients nearing EoL in PR; 2) Examine the perceived barriers to EoL discussion and decision-making process among oncologists in PR; and 3) Determine the knowledge, attitudes, and EoL planning among cancer patients and their caregivers in PR. This study will provide new evidence that will help us understand the complex process of EoL care and how different factors interact. Our most important contribution will be to document the potential multilevel influences on EoL care. This information will be essential for developing and implementing a multilevel intervention to improve the quality of care at the EoL. We expect our results can help improve health care delivery and enhance cancer patients' overall quality of life in a cost-effective way. | Research Centers | 6project_grants_public |
gen_fa61cf0bebfe15843beec0cada262213 | Sit-and-wait pathogens: Consequences of heterogeneity in pathogen exposure dynamics for environmentally persistent pathogens | NIH | VIRGINIA POLYTECHNIC INST AND ST UNIV | 1R01GM152978-01 | Transmission is a fundamental component of host-pathogen systems. Despite complex biological processes interacting to determine whether a pathogen successfully infects a host, transmission parameters often assume a constant per-contact transmission probability. Factors that can influence risk of infection include the frequency of exposures, duration of a contact, and dose acquired during an interaction, which are collectively referred to as transmission determinants. These determinants of infection can vary among hosts and between routes of transmission (e.g. direct and indirect) and have profound effects on disease dynamics. Ultimately, a mechanistic understanding of how heterogeneity in transmission probabilities among exposure events contributes to disease dynamics remains an important outstanding question in the ecology and evolution of infectious diseases. Here we seek to elucidate how factors determining transmission success influence the probability of acquiring infection, epidemic dynamics, and pathogen evolution. Specifically, we will investigate how transmission determinants (pathogen dose, contact duration, and contact frequency) vary among exposure routes (direct and indirect) and ultimately contribute to the dynamics of an emerging multi-host disease of snakes. Snake fungal disease (SFD) is caused by, Ophidiomyces ophidiicola, which is an environmentally persistent fungal pathogen that, to date, has been documented in more than 42 species of wild snakes on three continents, and has contributed to the severe declines of several species. O. ophidiicola is transmitted through direct and indirect contacts, and the behavior of snake species affected by SFD naturally vary along an exposure duration and contact intensity gradient, making this an ideal system for understanding the effects of variation in exposure events on disease dynamics. This work will highlight important mechanisms that contribute to variation in snake declines, and more broadly, provide insight into the theoretical underpinnings and profound effects that factors determining successful transmission can have on infectious disease outbreaks. This proposal will leverage our recent advances in this multi-host system and use a combination of field, experimental, and modeling approaches to provide critical insight into how the dynamics of exposure events have cascading effects on infectious disease. Our research will provide a framework for understanding the role of transmission determinates in infectious disease dynamics across a broad array of infectious disease systems. Our work will therefore contribute to the control of infectious diseases to enhance human health and reduce illness. | Non-SBIR/STTR | 6project_grants_public |
gen_3a3f343be1a6a6c91b1193c717fd1a5d | Pathogenesis of Restrictive Allograft Syndrome Post-Lung Transplantation | NIH | EMORY UNIVERSITY | 7R01HL162171-03 | PROJECT SUMMARY/ABSTRACT Restrictive allograft syndrome (RAS) is a particularly aggressive form of chronic rejection post-lung transplantation, marked by an extremely poor prognosis (median survival of < 1 year) and little therapeutic options. Pathogenesis of RAS remains elusive although association with donor specific antibodies and antibody mediated rejection have been found in human studies. We have recently characterized a murine lung transplant model which recapitulates the histologic changes of RAS and establishes the requisite role of humoral immune response in its development. A unique feature noted in the murine RAS lung allograft was the presence of activated B cells, plasmablasts, and fully differentiated plasma cell (PCs). PCs were also identified in human RAS lungs suggesting that a rejecting lung serves as an inflammatory local niche for humoral immune responses. These antibody secreting cells (ASCs) localized along the bronchovascular bundles (BVBs) and sub-pleural space, lying in close association with expanding mesenchymal cells (MCs). The key role of specialized stromal cells, via paracrine factors of C-X-C Motif Chemokine Ligand 12 (CXCL12) and interleukin (IL)-6, in establishment of a stable survival niche for ASCs is well recognized across a variety of lymphoid and hematopoietic tissues. We have recently characterized the in situ niche of a subpopulation of lung-resident mesenchymal cells which expand and contribute to fibroproliferation in a RAS allograft. This Foxf1+/Gli1+/Scal1+/Col1+ mesenchymal stromal cell (MSC) population forms a three dimensional network along the bronchovascular bundle (BVB-MSCs). Our new preliminary data demonstrates that these cells are the high CXCL12/IL-6 expressing population, and lie in close apposition to ASCs in the rejecting lung allograft. A novel mechanism of upregulation of CXCL12 expression in BVB-MSCs by IL-6 transsignaling and downstream JAK/Stat activation, with recruited monocytes contributing to the soluble IL-6R, was identified. In this proposal we will investigate an innovative and novel hypothesis of paracrine signaling between this graft-resident mesenchymal stromal cell population and immune cells and the role of IL-6 transsignaling/CXCL12 axis in regulating the humoral inflammatory niche in a rejecting lung. The proposed experiments will utilize our ability to identify and conditionally target the specific BVB-MSC subpopulation, and the novel orthotropic whole lung transplant model of RAS to elucidate the spatial in vivo niche of APCs, its temporal regulation by CXCl12 expressing BVB-MSCs, and its functional significance in the pathogenesis of RAS (Aim 1). The role of IL-6 and IL-6 trans-signaling in cellular communication between resident MCs and infiltrating immune cells within this niche and the pathogenesis of RAS will be determined (Aim 2). The effect of specific drugs targeting humoral cell responses and IL-6 signaling pathway will be tested in the murine lung allograft model of RAS (Aim 3). Together these studies will offer novel mechanistic insight into lung allograft failure and provide pre-clinical information regarding role of therapeutic modalities targeting humoral immune responses in RAS. PROJECT NARRATIVE Lung transplantation is a costly and life-saving procedure with the drawback of poor long-term survival secondary to development of chronic rejection. In this application, we will investigate the mechanism and potential therapeutic targets for a particular aggressive form of chronic rejection. | Non-SBIR/STTR | 6project_grants_public |
gen_e8ab680233030fd63dcd694b882e7bda | Innate immune regulation of wound re-epithelialization | NIH | UT SOUTHWESTERN MEDICAL CENTER | 5R00AR072780-04 | PROJECT SUMMARY/ABSTRACT Skin has a remarkable ability to heal wounds through re-epithelialization, a repair process fueled by adult stem cells residing in the epidermis and hair follicles. Following injury, wound-edge keratinocytes proliferate and migrate to initiate wound closure, which is accompanied by activation and infiltration of immune cells. My long-term goal is to elucidate the cellular and molecular basis underlying wound re-epithelialization, how the immune system regulates this process, and how it affects tissue regeneration. Previously we found activation of the transcription factor Stat3 in keratinocytes controls many important aspects of wound re-epithelialization, including basal keratinocyte proliferation, migration and crosstalk with epidermal dendritic T cells (DETCs). However, the molecular mechanism by which wounding triggers Stat3-mediated re-epithelialization and activates the immune system remains unclear and is the subject of this study. Cellular injury is known to produce damage associated molecular patterns (DAMPs) that are sensed by the innate immune system for host protection. We hypothesize that DAMPs produced by skin wounds are sensed by innate immune pattern recognition receptors (PRRs), which then signal to produce cytokines, and further activate Stat3 for wound re-epithelialization. Using a candidate approach and Stat3 activation as a readout, we will first identify, characterize, and verify wound-edge cytokines that influence wound re-epithelialization through epidermal-specific genetic knockouts, gene-expression analysis, and genetic modulation of immune signaling (Aim 1). Next, we describe strategies to identify the immune signaling pathway, upstream PRR, and the cells responsible for the PRR signaling through genetic and biochemical approaches (Aim 2). Finally, we describe an inducible genetic model of wound injury, characterize its similarity to physical wounding, and identify wound- induced ligands using biochemical purification and an in vitro assay (Aim 3). These lines of investigation will 1) offer novel insights into the molecular mechanism of wound initiation and innate immune contribution to skin re-epithelialization, 2) contribute new tools and models to the study of immune regulation and skin repair, and 3) improve our understanding and therapeutic options for autoimmune/autoinflammatory skin conditions and diseases associated with poor wound repair. With an exceptional mentoring team led by Dr. Elaine Fuchs (with Drs. Jean-Laurent Casanova and Daniel Mucida) and a supportive, stimulating training environment at the Rockefeller University, I am ideally positioned to fully develop my technical skills and knowledge in skin biology and immunology. My research, training, and career development will allow me to establish a unique niche in the field of wound-repair and tissue regeneration as an independent investigator. PROJECT NARRATIVE The characterization of innate immune regulation of skin wound-repair will provide insights into the pathology and therapeutic options for autoimmune/autoinflammatory skin conditions such as psoriasis and atopic dermatitis. The identification of molecular mechanisms that trigger skin-wound repair will contribute to our understanding and treatment for conditions associated with chronic non-healing wounds such as diabetic ulcers, epithelial cancer, and poor wound-repair associated with aging. | Non-SBIR/STTR | 6project_grants_public |
gen_08c5bf95eac90725e5344f4b4c8d4a44 | Understanding Chirality at Cell-Cell Junctions With Microscale Platforms | NIH | RENSSELAER POLYTECHNIC INSTITUTE | 1R01GM141387-01A1 | The regulation of cell-cell junctions is essential for the biological functions of various tissues such as epithelium and endothelium. Recent evidence in embryonic development and vascular physiology suggests that cell-cell junctions are regulated by cell chirality, a universal but fundamental property of the cell. We pioneer in research in cell chirality using engineered in vitro platforms. Here, using these platforms, we are to investigate the biophysical mechanism associated with chirality at cell-cell junctions. While the principle may be shared among many cell types, this study will focus on endothelial cells and the regulation of vascular permeability. The endothelial cell layer is a semi-permeable barrier that tightly controls the passage of proteins and cells in the bloodstream into the interstitial space and regulates the local environment of biological tissues in living organisms. Cells achieve this vital function primarily through mediating paracellular transport by controlling the opening and closure of cell-cell junctions. Protein Kinase C (PKC) activation has been associated with endothelial dysfunction in chronic conditions such as diabetes and long-term smoking as well as acute diseases such as sepsis, acute lung injury, and viral infection. Restoring and maintaining vascular integrity is critical for body function and patient survival, especially for acute diseases. Recently, we have demonstrated that PKC can reverse cell chirality, which mediates endothelial permeability. However, little is known about the molecular mechanism of how PKC activation reverses endothelial chirality or that of how cell chirality alters endothelial permeability. In this proposal, we hypothesize that PKC reverses cell chirality by reducing the level of actin crosslinking and that cell chirality regulates cell-cell junctions (and therefore endothelial permeability) biomechanically through actin tilting and VE-cadherin localization. We will pursue the following three aims: Aim 1. Identify the timing and location of biomechanical asymmetry responsible for multicellular chiral morphogenesis using traction force microscopy (TFM). Combing 2D micropatterning for cell chirality and TFM for cellular forces, we are to study in great detail of 2D collective symmetry breaking and to interrogate underlying cellular biomechanical mechanisms. Aim 2. Determine cytoskeletal mechanisms underlying PKC induced reversal of endothelial cell chirality. We will identify formin isoforms and actin crosslinkers involved in this process, and their regulation by PKC signaling. Aim 3. Investigate the role of chirality mismatch in the intercellular gap formation and endothelial permeability. We will quantify actin structure and dynamics during the intercellular junctions and examine how the mismatch of cell chirality can lead to actin remodeling and induce intercellular gap formation. If successful, we will be able to identify the biophysical mechanisms, allowing for the potential development of novel, specific therapies based on cell chirality for endothelial dysfunction. With data obtained from this proposal, we will seek further support and examine our findings with animal models. Project Narrative Cell-cell junctions are tightly controlled during physiological activities. Vascular disorders change the leakiness of blood vessels drastically, leading to severe consequences including death. Here we propose to study the biophysical mechanism of cell handedness in the maintenance of vascular integrity. | Non-SBIR/STTR | 6project_grants_public |
gen_49583e64d7c7a300530c38863ab3d768 | Administrative and Biostatistics Core | NIH | SALK INSTITUTE FOR BIOLOGICAL STUDIES | 5P01CA265762-02 | PROJECT SUMMARY – Administrative and Biostatistics Core The overall goal of the Administrative and Biostatistics Core (Admin Core) is to facilitate research efforts toward understanding intrinsic resistance to therapy in pancreatic ductal adenocarcinoma (PDA). This will be accomplished by providing the three Research Projects (Projects) and the three Shared Resource Cores (Cores) with organizational support, professional project management, biostatistical expertise, data storage and sharing solutions, and a governance framework. The Admin Core will plan and coordinate meetings between the scientific teams, as well as between the scientific teams and the leadership groups. The Admin Core will include a Project Manager, who will provide professional project management services to ensure that stated objectives are successfully achieved. As part of this effort, the Project Manager will monitor Core activities to ensure that efforts of these cutting-edge scientific resources are integrated and coordinated, and that the Cores are operating effectively and efficiently to meet the needs of the Projects. The Project Manager will also establish a governance framework for resolving potential conflicts between team members and for managing resource utilization and budgetary concerns. The Admin Core will include a Biostatistics Unit that will be intimately involved in all experimental design and data analysis. The Biostatistics Unit will be responsible for ensuring that all experiments are sufficiently powered, and that the resulting data are analyzed using the most scientifically rigorous approaches. Novel methods will be developed when needed. One faculty-level statistician will split their effort between the Biostatics Unit and the bioinformatics team within the Tumor and Microenvironment Heterogeneity Core, coordinating communication and the transfer of data between these data analytic groups. Finally, the Admin Core will include a Data Curator who will help establish and oversee a centralized data-storage system into which all data generated by the scientific teams will be deposited and properly annotated. This resource will facilitate the integration of disparate datasets, as well as the exchange of data and reagents between the Projects, the Cores, and the external research community, as appropriate. Thus, the specific aims of the Admin Core are to: 1) provide overall organizational support including the coordination of meetings between the Projects, the Cores, and the leadership committees, 2) provide project management services to ensure that scientific objectives are achieved in an efficient and cost-effective manner, 3) provide biostatistical expertise to ensure experimental rigor and reproducibility, and 4) create and manage a centralized data storage and sharing system to enable data integration. PROJECT NARRATIVE – Administrative and Biostatistics Core Pancreatic ductal adenocarcinoma is a highly lethal malignancy that will soon become the second-leading cause of cancer-related death, in large part because patients often exhibit inherent and/or adaptive resistance to therapy. This program project will explore the epigenetic programs, signaling pathways, and metabolic adaptations that drive therapeutic resistance, with the end goal of revealing new therapeutic targets. The Administrative and Biostatistics Core will provide the leadership framework, communication pipelines, project management services, biostatistical expertise, and data storage/sharing solutions necessary to coordinate the activities of the three Research Projects and three Shared Resource Cores that comprise this P01 effort, thereby providing an organizational foundation for success. | Non-SBIR/STTR | 6project_grants_public |
gen_d985cdeac35e43129af03bcbd8ef5e5f | Improving vaccination against type-2 T-independent antigens | NIH | ROSALIND FRANKLIN UNIV OF MEDICINE & SCI | 5R21AI176124-02 | Summary Infections with encapsulated bacteria are responsible for the death of millions of children and the elderly each year. Capsular polysaccharides behave as T-independent antigens (TI) and vaccination based on these carbohydrates provides protection against these infections. The efficacy of vaccines against TI antigens is severely limited by their inability to induce recall responses upon revaccination or infection. This phenomenon depends on inhibition of memory B cells by antigen-specific IgG generated in the primary response. The mechanism behind this inhibitory activity however, remains unclear. The efficacy of vaccines critically depends on adjuvants that can promote strong memory and influence Ig class switch recombination (CSR). However, effective adjuvants for TI antigens vaccination are not available. The goal of our proposal is to answer two outstanding questions regarding the response to TI antigens: what is the mechanism of the inhibition of recall responses? and how can CSR and generation of memory B cells can be manipulated to improve vaccination? For these studies we will use a mouse model of tularemia and will use Francisella tularensis LPS, a model TI antigen that we showed activates B1 cells. In aim 1 we will test the hypothesis that IgG-LPS immune complexes inhibit memory B1 cells reactivation and IgM production through complement activation. We will test whether complement depletion or absence could restore the recall response to vaccination with TI antigens and result in a more effective vaccine for tularemia. In aim 2 we will test the hypothesis that novel adjuvants can improve recall responses to TI antigen by manipulating CSR in B1 cells and/or improving generation of memory B1. Based on our preliminary results we will test a number of potential adjuvant treatments with the goal of achieving sustained IgM and increased LPS-specific IgA production but reduced IgG (responsible for inhibition of recall response). These adjuvants are also expected to increased numbers of memory B1 cells. If successful, our studies will create proof of concepts that can inform future development of more effective vaccination strategies for TI antigens. Narrative Infections with encapsulated bacteria are responsible for the death of millions of children and the elderly each year. The efficacy of vaccines against these pathogens is severely limited by their inability to induce recall responses upon revaccination or infection. We will use a mouse model to improve vaccination against one of these bacterial infections and create proof of concepts that can inform future development of more effective vaccination strategies. | Non-SBIR/STTR | 6project_grants_public |
gen_ac8a0c91447bec0f275d07f37255f48d | The regulation of phosphoprotein phosphatases in the nucleus | NIH | UNIVERSITY OF CONNECTICUT SCH OF MED/DNT | 5R01GM144379-02 | ABSTRACT An estimated 70% of all eukaryotic cellular proteins are regulated by phosphorylation. Strict temporal and spatial control are essential for the fidelity of this process, as derailed signaling cascades lead to disease. While the importance of phosphorylation is clear, knowledge gaps remain in the mechanisms that regulate key proteins involved in this process, especially phosphoprotein phosphatases (PPP). Our long-term goal is to understand the structural and functional mechanisms that control PPP activity in health and disease. Here, we focus on the function of protein phosphatase 1 (PP1) and PP2A, both of which have major roles in cell division and cancer. Our aims are designed to define the mechanisms of PP1- and PP2A:B55-based substrate recruitment to obtain a systems biology understanding of the proteomes and phosphatomes directed by these enzymes. For the PP2A family of enzymes, it is established that substrates are recruited by their variable B- subunits. We recently showed that the PP2A B56 subunit binds specifically to its substrates via a newly identified short linear motif (SLiM), LpSPIxE. This has led to the discovery of scores of novel B56-specific substrates and the development of the first PP2A:B56-specific regulator. Here, we investigate PP2A:B55, the most abundant PP2A holoenzyme in cells and the primary enzyme responsible for dephosphorylating CDK1 targets to initiate mitotic exit. Consistent with this, at mitotic entry, PP2A:B55 activity is inhibited. This is achieved by two B55-specific inhibitors: FAM122A and ARPP19. To molecularly define how these inhibitors block PP2A:B55 activity and to elucidate the molecular basis of B55 substrate recruitment via a B55-specific SLiM, we will determine both holoenzyme (quadruple complexes) structures. This is technically challenging, as these PPPs cannot be functionally expressed in E. coli or insect cells, a problem we have successfully overcome. Furthermore, we have developed a unique PP1 regulator (PhosTAP), which we show can be successfully leveraged to fully define the PP1 interactome and phosphatome. Due to its 100% specificity and exceptional affinity for only PP1, this novel PP1 PhosTAP can also be leveraged to specifically recruit PP1 to its point of action within the cell, in a manner similar to that used by PROTACs for targeted degradation. Together, the proposed aims will provide the much-needed molecular data that demonstrate how key PPP holoenzymes, especially PP1 and PP2A holoenzymes, bind their substrates and how these interactions are regulated during the cell cycle. Because these holoenzymes have critical roles in multiple human diseases, especially cancer, the proposed work will establish these holoenzymes specifically, and PPPs generally, as potent and specific drug targets. Project Narrative: The protein phosphatases PP1 and PP2A are ubiquitous ser/thr protein phosphatases that are critical during mitosis. Despite decades of research, most PP1 and PP2A substrates are still unknown. We have assembled a team of researchers that will overcome this challenge to allow for novel biological insights to be gained. | Non-SBIR/STTR | 6project_grants_public |
gen_bba59e927cd5ed115c8aae8cbfc98258 | Modeling the dynamics and composition of T-cell receptor repertoires in post-acute sequelae of COVID-19 (PASC) | NIH | INSTITUTE FOR SYSTEMS BIOLOGY | 1R03AI175977-01A1 | Project Summary Post-acute sequelae of COVID-19 (PASC), colloquially known as long covid, has been reported in 31%-69% of COVID-19 patients. The prevalence of PASC among patients, even those with mild infection, and those with prior vaccination, underscores the importance of principled approaches for uncovering and addressing the cause of these issues. We propose to conduct a detailed analysis of the immune repertoire dynamics of PASC patients and compare them to both COVID-19 patients without PASC, and healthy individuals. These will be secondary analyses, conducted on existing data from a previous study with the Institute of Systems Biology and Swedish Health Services, which includes longitudinal deep immunophenotyping with single cell and plasma multiomics, repertoire sequencing, electronic health records, viremia measurements, and antibody titers for 209 COVID-19 patients. In Aim 1, we will conduct inference, analysis, and comparison of T-cell receptor repertoire dynamics in PASC. Using interpretable statistical, biophysical, and machine learning approaches, we will conduct a detailed analysis of T-cell repertoires aimed at finding PASC specific clonotypes and their corresponding receptor features. This involves building cohort specific models of thymic selection, examining how these models differ between cohorts and over time, inferring the dynamics of repertoire size, sharing and diversity, and uncovering the receptor features which drive these differences. In Aim 2, we will develop new methods for the integration of T-cell repertoire and single cell dynamics. The existing breadth of multiomics data allows us to explore new methodologies for integrating different modalities of longitudinal data. For T-cell repertoires in particular, we plan to extend existing models of thymic selection to include gene expression of relevant T-cell specific genes and to study how the expansion and contraction of clonotypes affects the dynamics of T-cells in gene expression space. By using interpretable biophysical and machine learning methods, we can construct generative models of TCRs including gene expression values and study how these distributions change in time. Results have strong potential to accelerate our understanding of the etiology of immune-based PASC responses, which is essential for prioritizing potential therapeutic targets for prevention and treatment. Further, results will advance methods for future research across a wide range of infectious diseases and immune-mediated medical conditions. Narrative Using secondary data from previous studies, we will conduct a detailed analysis of immune repertoire dynamics in patients with post-acute sequelae of COVID-19 (PASC), as compared with patients with a history of COVID-19 without PASC, and with healthy individuals. We will build cohort-specific models of thymic selection, examining how these models differ between cohorts and over time, inferring the dynamics of repertoire size, sharing and diversity, and uncovering the receptor features which drive these differences. In addition, we will develop new methods for the integration of T-cell repertoire and single cell dynamics, which will be of benefit for future research across a wide range of infectious diseases and immune-mediated medical conditions. | Non-SBIR/STTR | 6project_grants_public |
gen_5cbf93ae6efc758ca80be0648af07528 | Assessing the Impact of SARS-CoV-2 on Adipose Tissue Function and Glucose Homeostasis | NIH | WEILL MEDICAL COLL OF CORNELL UNIV | 3R01DK132879-01A1S1 | Project Summary/Abstract COVID-19 has proven to be a metabolic disease resulting in adverse outcomes disproportionally afflicting individuals with diabetes or obesity. Patients infected with SARS-CoV-2 and hyperglycemia suffer from longer hospital stays, increased need for mechanical ventilation and mortality compared to those without hyperglycemia. We found that insulin resistance rather than beta cell failure is the predominant cause of hyperglycemia in acute COVID-19. The insulin sensitizing hormone adiponectin is diminished in the circulation of COVID-19 patients compared to controls. Furthermore, we demonstrate that SARS-CoV-2 can directly infect adipocytes. Importantly, we find replicating virus in adipose tissues of both autopsy samples from COVID-19 patients and in mouse and hamster experimental models of SARS-CoV-2 infection. Together these data suggest that SARS-CoV-2 triggers adipose tissue dysfunction to drive insulin resistance and adverse outcomes in acute COVID-19. In this proposal, we seek to follow up on these studies and assess the mechanisms driving adipose tissue dysfunction in acute and recovered models of COVID-19. We will pursue the following specific aims: 1. Assess the impact of acute SARS-CoV-2 infection on glucose homeostasis in obese and non-obese mice. 2. Map the spatiomolecular interactions and dissect the molecular mechanisms of SARS-CoV-2 infection in adipose. 3. Determine the long-term glycometabolic consequences of SARS-CoV-2 infection. The overall goal of these studies is to assess how COVID-19 can drive adipose tissue dysfunction and hyperglycemia and will shed light on novel targets to combat metabolic complications induced by COVID-19. Public Health Relevance The deadly COVID-19 pandemic is underscored by the high morbidity and mortality rates seen in certain vulnerable populations, including patients with diabetes mellitus (DM) and obesity. Despite this well-known association, our understanding of how and why this occurs is lacking leading to an absence of specific therapies. In this proposal, we aim to test the mechanisms by which COVID-19 leads to adverse outcomes in diabetes mellitus and obesity with the goal of developing new therapies for patients with COVID-19. | Non-SBIR/STTR | 6project_grants_public |
gen_78f0d6789225ecaa56fdabeba3d41865 | An in vivo multiplex model to study gene-environment interaction in Parkinson's Disease | NIH | UNIVERSITY OF ROCHESTER | 3R00ES033723-03S1 | Project Summary/Abstract Parkinson's disease (PD) is a progressive neurodegenerative disorder that is characterized by α-synuclein-rich neuronal inclusions. Recent genome-wide associated studies (GWAS) and epidemiological studies have identified multiple candidate genes and environmental factors which can modify PD risk. However, studying polygenic interactions with environmental factors has been difficult due to the lack of a model system. However, studies have hinted at a complex relationship between α-synuclein, genetic risk factors, and environmental factors. In our preliminary data, we have established a multiplex model using the Drosophila model of PD. In this model, we express human α-synuclein, simultaneously modify GWAS candidate genes in neurons, and expose adult flies to rotenone. Using a combination of scalable techniques in this model, we identified novel interactions among α-synuclein, environmental factors, and GWAS genes. The overarching hypothesis is a multiplex model, in combination with iPSC-derived neurons, can be used to identify and study the mechanism of novel gene-environment interactions. Further, this model system will identify potential drug targets that can modify the gene-environment interactions. In Aim 1, a series of experiments, including super-resolution microscopy and iPSC-derived tyrosine hydroxylase (TH) neurons, will be performed to characterize the interaction among LRRK2, rotenone, and α-synuclein, which was identified using the multiplex model. Aim 2 will involve understanding the mechanism of interactions among LRRK2, rotenone, and α-synuclein. Previous studies and preliminary experiments have shown that actin hyperstabilization plays a central role in regulating neurotoxicity. Herein biochemical, immunohistological, and neurotoxicity assays will be performed in Drosophila and iPSC-derived TH neurons (obtained disease-causing LRRK2-G2019S and protective LRRK2-R1398H iPSCs) to study the role of actin dynamics in regulating this gene-environment interaction. Aim III will identify a druggable target that can modify the interaction among LRRK2, rotenone, and α-synuclein. Further, we will screen for other PD-related neurotoxicants that interact with LRRK2 and α-synuclein through actin hyperstabilization. Finally, we will genetically and pharmacologically inhibit MRCKα, a kinase that can regulate actin hyperstabilization, in flies, iPSC-derived neurons, and a mouse model. This project may elucidate a novel model system that can be used to identify and study the mechanism of gene- environment interactions. My training during the K99 phase enabled me to transition to an independent position at URMC. I will lead a laboratory investigating the molecular mechanisms of gene-environment interactions in neurodegenerative disorders. Public Health Relevance Statement: Project Narrative Due to the lack of model systems, it has been difficult to study complex gene-environment interactions in Parkinson's disease. Using a combination of Drosophila genetics and patient-derived induced pluripotent stem cells, we can develop a model that can be utilized to identify and study the mechanisms of interaction among GWAS candidate genes, environmental factors, and α-synuclein. This K99/R00 proposal aims to characterize a novel interaction among LRRK2, rotenone, and α-synuclein, study the mechanism of the interaction, and subsequently validate this novel multiplex system, which can be used to study these previously unknown interactions among environmental and genetic factors and is a step towards personalized medicine. | Non-SBIR/STTR | 6project_grants_public |
gen_9bd58a99e4d21d1c355371c9a381446c | Composing CODAs to cervical cancer screening through an integrated CRISPR and fluorescent nucleic acid approach | NIH | MASSACHUSETTS GENERAL HOSPITAL | 5U01CA279858-02 | ABSTRACT Challenges. Despite being curable when caught early, cervical cancer remains the second leading cause of death in women living in sub-Saharan Africa. Even with available screening technologies (cytology, visual inspection with acetic acid (VIA), and sometimes high-risk Human Papillomavirus [hrHPV] testing), only 10-20% of eligible women in low-and-middle-income countries (LMIC) are currently screened. Barriers cited include technical expertise, laboratory capacity, cost, and access. Most developed countries have adopted PCR-based HPV screening, furthering the resource divide with LMICs, where 80% of cancer deaths occur. Innovation. In dire response to the diagnostic gaps unveiled by the pandemic, team members within our group developed and validated in human specimens a robust and frugal technology to detect nucleic acid targets. The approach, CRISPR Optical Detection of Anisotropy or CODA, combines CRISPR/Cas (a Nobel Prize- winning technology for highly precise gene editing) with fluorescence anisotropy (differential rotational motion of fluorescent molecules). Automated nucleic acid readouts are generated in < 30 minutes. We have adopted this technology for: a) the detection of DNA and RNA markers of CIN2+; and b) reliable use by clinicians and clinical lab personnel in point-of-care settings. Our plan for this proposal is to operationalize this technology for low resource settings, notably our partner sites in Uganda and Ghana, with close LMIC input. We will then leverage CODA and other parameters to create a panel of tests to optimally detect CIN2+ in a single encounter. Aim 1: Construct a robust CODA platform for comprehensive HPV screening. Aim 2: Further examine CODA performance on human specimens and refine for LMIC operations. Aim 3: Develop a novel and rapid multi-modal algorithm for screen and treat in LMICs. Impact. Given CODA's core strengths in nucleic acid analyses (e.g., DNA or RNA), we envision an approach that yields fully quantitive readouts of high-risk HPV DNA and E6, E7, and p16 mRNA. Since the CODA assay underlies all readouts driving this proposal, a high potential exists for end-user-friendly, practical, and rapid triage of high-risk cervical disease or invasive cancer. These benefits could help decentralize and harmonize screening efforts with those guidelines currently endorsed by resource-rich countries. NARRATIVE Cervical cancer, while curable when caught earlier, remains a leading cause of death in low-and-middle- income countries. Inadequate screening tactics arise from various factors including pathology bottlenecks, costs, clinical bandwidth, and turnaround times. We propose to advance a low-cost and fast portable platform (based on Nobel Prize-winning technology) for point-of-care and integrated cervical cancer screening in Uganda and Ghana. If successful, cervical cancers would be identified earlier through a sustainable, decentralized approach. | Non-SBIR/STTR | 6project_grants_public |
gen_2956ae1404c7a8b9f59c4c2c49f27761 | Interrogating function, regulation, and interactions in a clade of prevalent human gut microbes | NIH | OHIO STATE UNIVERSITY | 5R35GM151155-02 | PROJECT SUMMARY/ABSTRACT Human-associated microbes are causally linked to processes as diverse as immunomodulation, protection against pathogens, and atherosclerosis. Many of these links are mediated through biochemical transformations of dietary, drug, or host compounds. This makes the microbiome a promising therapeutic target, especially as we could potentially affect downstream processes by controlling metabolic inputs. However, in order to effectively intervene, we must first understand how exactly changes in these inputs lead to differential regulation of growth, gene expression, and metabolism. This is challenging because our microbiomes are not only genetically and physiologically diverse, but are also highly diverged from the most common model organisms, with many genes of unknown function. Over the next five years, my research group will use a combined computational and experimental strategy to characterize gene function, metabolic regulation, and microbial interactions in one of the most prevalent and abundant clades of gut bacteria, the Bacteroidales. Specifically, we seek to determine 1) which Bacteroidales genes are involved in growth on different nutrients and stressors; 2) how Bacteroidales genes are regulated, and how this affects their metabolic outputs; and 3) how Bacteroidales interact with the other microbial inhabitants of the gut. We will accomplish this by gathering high-throughput in vitro data from diverse sets of microbiome isolates and synthetic communities, developing more powerful and specific statistical tools to analyze these data, and using these new data and tools to re-analyze metagenomics data gathered from in vivo case-control studies. I envision that this line of inquiry will provide missing fundamental knowledge about this clade of microbes, which will ultimately help us interpret case-control studies of the microbiome and support the development of more precise interventions. PROJECT NARRATIVE Our gut microbes have many well-established effects on our health, but many of the most common gut microbes are only distantly related to typical laboratory model organisms. The proposed research develops quantitative, data-rich approaches to understand how these microbes grow, behave, and interact in different environments. In the future, this line of research could both improve our ability to interpret case-control studies of the microbiome, and lead to more precise, safer therapies that target our microbiome to improve health. | Non-SBIR/STTR | 6project_grants_public |
gen_facb5416f7ba2ad509fba54bb4204df3 | In vivo Evaluation of Lymph Nodes Using Quantitative Ultrasound | NIH | WEILL MEDICAL COLL OF CORNELL UNIV | 5R01CA277038-02 | Project Summary This application proposes a collaborative study to be performed by Weill Cornell Medicine (WCM) in New York, NY, as the lead organization in collaboration with GE Research (GER) in Niskayuna, NY, and Stony Brook Medicine (SBM) affiliated with the State University of New York in Stony Brook, NY. The proposed project addresses the need for reliable, highly sensitive means of detecting metastases to lymph nodes (LNs) and distinguishing them from primary lymphomas and LNs affected by benign conditions. This capability will allow improved staging and treatment of disease. Accordingly, we seek to validate existing encouraging results obtained in prior studies by WCM and SBM using quantitative-ultrasound (QUS) methods to detect metastases in LNs by applying and evaluating these promising methods using a modified clinical scanner to acquire ultrasonic echo-signal data from patients undergoing medically required ultrasonically guided fine-needle aspirations (FNAs) of suspicious LNs. The study will include a far broader range of disease types from a larger and more-diverse patient population than has been possible in studies to date. Validation efforts will develop, deploy, and validate an application for QUS- based detection and characterization of LNs implemented in a clinical scanner. Successful validation will establish a foundation for incorporation of QUS-based methods for LN evaluation into clinical ultrasonic scanners. Such instruments will be able to evaluate suspicious LNs intra-operatively in the operating room or pre-operatively in the examination room. The collaborating researchers will synergize capabilities in the following general ways: WCM will provide project oversight and coordination and will apply, refine, and test QUS and classification methods; GER will provide ultrasound instrumentation and will develop and validate the QUS application for the scanner; SBM will recruit patients and acquire ultrasonic and cytopathological data from patients undergoing FNAs of their LNs. Narrative Reliable evaluation of lymph nodes (LNs) is essential for proper disease management. However, current non- invasive methods cannot reliably distinguish metastatic LNs from non-metastatic LNs. The proposed study will develop low-cost, in-vivo, ultrasonic instrumentation for more-reliable metastasis detection and hence for improved staging and treatment of disease. | Non-SBIR/STTR | 6project_grants_public |
gen_734bd2af99b4b8c313c924535b9fadaa | Endothelial Metabolic Reprogramming by Interferon-gamma in Coronary Artery Disease | NIH | BRIGHAM AND WOMEN'S HOSPITAL | 5K08HL165047-02 | PROJECT SUMARY/ABSTRACT This proposal presents a five-year research and training program to establish Laurel Y. Lee, M.D., D.Phil. as an independent, R01-funded physician-scientist in academic cardiology with expertise in immune modulation of endothelial metabolism in atherosclerosis. This unique scientific focus combines Dr. Lee’s doctoral training in T- cell immunology with her subsequent clinical and research fellowships in cardiovascular medicine at the Brigham and Women’s Hospital (BWH) and Harvard Medical School (HMS). She is currently an Associate Physician in the Division of Cardiovascular Medicine and an Instructor in Medicine at BWH/HMS. Coronary artery disease remains a leading cause of mortality and morbidity worldwide. While endothelial dysfunction is known as a precursor to atherosclerosis, how altered endothelial metabolism contributes to atherogenesis remains incompletely understood. The principal investigator’s long-term goal is to define how local immune activation alters endothelial metabolism and contributes to atherogenesis. As a first step toward achieving this goal, she recently discovered that interferon gamma (IFN-γ), a T-cell cytokine abundant in human atheroma, impairs endothelial glucose metabolism and activates fatty acid oxidation in primary human coronary artery endothelial cells (Lee et al., Circulation, 2021). These metabolic derangements were associated with proatherogenic endothelial phenotypic changes, raising the central hypothesis that IFN-γ-induced endothelial metabolic reprogramming forms a novel mechanistic basis for accelerated atherosclerosis. This hypothesis will be tested through the following aims: (1) Define the effect of IFN-γ on endothelial fuel utilization, (2) Establish the mechanistic link between endothelial metabolic reprogramming and endothelial phenotypic changes, and (3) Define the changes in endothelial metabolism in a mouse model of immune exacerbated atherosclerosis in vivo. Using the cutting-edge approaches including metabolomics, vascular phenotyping, single-cell technology, and a mouse model of atherosclerosis, the principal investigator will acquire new skills and expertise in quantitative analyses of metabolism, lipid biology, and in vivo analysis of immune-endothelial interaction in experimental atherosclerosis. These studies, if successful, will establish immune mediated endothelial metabolic perturbations as a novel mechanistic basis for linking pathologic T-cell activation and atherosclerosis and may open new therapeutic strategies. Dr. Joseph Loscalzo, a distinguished vascular biologist with expertise in vascular metabolism, redox biochemistry, and systems biology will serve as the principal investigator’s primary research mentor. An advisory committee of physician-scientist experts in cellular metabolism and atherosclerosis research will provide further scientific and professional development guidance and assessment of her progress. In summary, Dr. Lee has created a superb environment and mentoring team to develop her unique niche in immune modulation of endothelial metabolism. The proposed research, training plans, and outstanding environment at BWH, HMS, and MIT will propel her transition to an independent investigator and a leader in vascular research. PROJECT NARRATIVE This proposal will define how interferon-gamma, a major T-cell cytokine abundant in atherosclerotic plaques, alters fuel utilization by the vascular cells that line the innermost layer of the coronary arteries, thereby making these vessels susceptible to atherosclerosis progression. This study, if successful, will improve our understanding of the adverse changes involving vascular metabolism in atherosclerosis and may lead to new therapeutic strategies to prevent heart attacks, strokes, and limb loss. | Other Research-Related | 6project_grants_public |
gen_5950eb47b0079bdc6ed85473006db1bd | Development and Evaluation of Advanced Non-Contrast Perfusion MRI for Monitoring Treatment Response in Brain Metastases | NIH | JOHNS HOPKINS UNIVERSITY | 1R01CA282928-01 | Project Abstract Brain metastases (BM) are the most commonly diagnosed type of central nervous system tumor, more frequent than primary intracranial neoplasms. Progress on various therapies have accelerated over the past decade through ongoing clinical trials, and the historically poor outcomes for patients with BM have been markedly improved. Contrast-enhanced T1-weighted MRI is routinely applied to depict BM with the size of the enhanced lesions for assessing treatment response. Lesion enhancement due to the disruption of blood-brain barrier is rather nonspecific of the functions of brain tumors. The most studied MRI methodology for perfusion measurement is dynamic susceptibility contrast perfusion weighted imaging (DSC-PWI), which measures cerebral blood flow (CBF) and cerebral blood volume (CBV). CBV is the widely adopted perfusion measure as a sensitive marker of tumor vascularity. However, its clinical applicability in BM studies is hampered by its lack of absolute quantification, the contrast-leakage effect, and frequent susceptibility artifacts. Arterial spin labeling (ASL) is ideal for frequent non-invasive longitudinal monitoring of tumor vascularity. The standardized spatially selective ASL technique for CBF mapping is the pseudo-continuous ASL (PCASL) method using a single post- labeling delay, which may render underestimation of CBF due to transit time delay caused by slow arterial flow typical in elderly patients. Velocity-selective ASL (VSASL) was proposed to remove the time-delay sensitivity. Our group has implemented the first velocity-selective inversion (VSI) based VSASL with 3D segmented GRASE acquisition and demonstrated its higher sensitivity to perfusion signal over conventional ASL methods. Additionally, our group first developed VSASL based CBV mapping by removing labeling delay, which delivered much higher SNR than ASL based CBF mapping. Furthermore, our preliminary data showed that VSASL with 3D stack-of-spiral based FLASH acquisition delivered better perfusion image quality with less artifacts than using GRASE, and high temporal resolution potentially allowing adequate retrospective motion correction. The purpose of this study is: Aim 1, to conduct further technical developments for VSASL based CBF and CBV mapping protocols with accelerated acquisitions; Aim 2, to evaluate the sensitivity of the two optimized VSASL protocols to CBF and CBV changes within a month after the radiation therapy, and assess their early prediction to treatment outcomes; Aim 3, to compare the specificity of VSASL derived CBF and CBV values in the distinction of metastatic recurrence from radiation-induced effects; Aim 4, to ensure high reproducibility of the VSASL protocols between multiple scanners with different vendors and field strength. By completing the proposed aims, the advanced VSASL based CBF and CBV mapping methods are expected to demonstrate important values for monitoring treatment response in BM, will be readily available for large-scale clinical studies, and can benefit for studies of all primary and metastatic tumors in the body. Project Narrative Brain metastases (BM) are the most commonly diagnosed type of central nervous system tumor. Arterial spin labeling (ASL) is ideal for frequent non-invasive longitudinal monitoring of tumor vascularity. This project proposes a prospective and comprehensive evaluation of state-of-art ASL perfusion mapping methods on BM patients for both early predicting treatment outcome and distinguishing true recurrence from radiation effects during the follow-up monitoring, which can be readily utilized as a practical and cost-effective imaging biomarker for multi-center and multi-vendor clinical studies of BM. | Non-SBIR/STTR | 6project_grants_public |
gen_50c47b069b83b653b987a4f795969c0d | Team Support to Improve Glycemic Control Using CGM in Diverse Populations (TEAM CGM) | NIH | UNIV OF MASSACHUSETTS MED SCH WORCESTER | 5R01DK133265-02 | PROJECT SUMMARY Continuous glucose monitoring (CGM) has demonstrable benefits for people living with diabetes, including improvement in diabetes control and reduction in hypoglycemia. Randomized controlled trials have demonstrated that CGM can reduce hemoglobin A1c (HbA1c) and increase in the time in range (TIR) metric. Little is known about CGM use in the broader population with type 2 diabetes (T2DM) in low-income, minority populations not receiving insulin therapy. This proposed study will rigorously evaluate CGM in a diverse and vulnerable population with T2DM in the primary care setting. The proposed study will integrate CGM into our previously studied approach of mobile health (mHealth) diabetes management. Our prior research has leveraged mHealth tools and a community health worker (CHW)/clinical pharmacist team to manage low- income, minority populations with T2DM. Clinical pharmacists embedded in the healthcare system review patient glucose levels, promote medication adherence and collaboratively adjust therapy to help patients reach HbA1c goals. CHWs augment pharmacist-led efforts and address social determinants of health and provide individualized, contextual self-management support. In partnership with Baystate Community Health Centers and UMass Memorial Family Medicine Health Centers, we propose an effectiveness study of team-supported CGM using a Sequential Multiple Assignment Randomized Trial (SMART) study design. We plan to study a diverse population with 318 T2DM patients receiving pharmacist- and CHW-supported CGM delivered through community health centers (CHCs) in Massachusetts. The Specific Aims include: (1) Conduct a randomized, controlled trial to evaluate the effectiveness of pharmacist-supported CGM in a diverse patient population with T2DM in the primary care setting. We hypothesize that pharmacist-supported CGM will result in improved HbA1c, CGM metrics, and other secondary outcomes (e.g., quality of life) at 6 months compared with pharmacist-only care; (2) Re-randomize patients not at HbA1c goal after 6 months to receive or not receive CHW support beyond pharmacist-supported CGM. We hypothesize that adding CHWs will result in improved HbA1c, CGM metrics, and other secondary outcomes at 12 months compared with pharmacist-supported CGM alone; (3) Evaluate reach, effectiveness, adoption, implementation, and maintenance using the RE-AIM framework; and (4) Determine total cost and cost-effectiveness of CGM and the supportive components (e.g., clinical pharmacist and CHW) from the perspective of the healthcare organization. If this team support model of diabetes care is found to be cost-effective, such evidence may influence insurance restrictions on ambulatory CGM coverage in T2DM. PROJECT NARRATIVE Continuous glucose monitoring (CGM) can help improve blood sugar management in type 2 diabetes. A sequential, multiple assignment, randomized trial will evaluate clinical pharmacists, community health workers, and telehealth in supporting CGM use to improve blood sugar control. The study will also investigate the translation of supported CGM into primary care and estimate both cost and cost-effectiveness. | Non-SBIR/STTR | 6project_grants_public |
gen_d797bc475ecbe97b23b95f600578e138 | Structural and functional basis of bacterial transcriptional regulation | NIH | LEHIGH UNIVERSITY | 1R35GM150644-01 | Project Summary / Abstract Antibiotic resistance poses an increasingly prevalent public health challenge, but advances in the development of new drugs has not progressed commensurately. A significant impediment lies in that many fundamental biochemical processes in bacteria remain poorly understood. The primary goal of the proposed research is to illuminate how bacteria respond and adapt to changes within their environments from a structural and mechanistic standpoint. Using the genetically tractable Gram positive organism Bacillus subtilis as our model system, we will focus our efforts on elucidating the molecular basis for copper-dependent transcriptional regulation and uncovering the drivers of enzyme specificity during the environmental stress response. Copper is required for bacterial survival, but excess levels of this transition metal can be toxic. Therefore, copper levels must be carefully controlled. The mechanisms by which copper export occurs have been extensively studied, but relatively little is known about copper import. We hypothesize that Cu uptake is regulated by Cu-dependent transcriptional repressors and the proteins under their control. Here, we will focus on a suite of proteins implicated in these processes, studying them at a detailed structural, molecular, and cellular level to better understand how they regulate copper uptake. In parallel, we will investigate what causes a family of very closely related kinases to specifically regulate distinct functions. Many bacterial species, including Bacillus subtilis and a number of pathogenic strains, encode one or more kinases that contain a domain termed the Bergerat fold. Despite their structural commonalities, enzymes with this domain exhibit strong preferences for their physiological binding partners. However, the molecular basis for how such specificity is conferred has yet to be investigated. We hypothesize that variations within the Bergerat fold influence substrate specificity and kinetics and can be targeted by small molecule inhibitors. By integrating tools from structural biology, microbiology, biochemistry, and computational studies, we will be poised to reveal new paradigms in transcriptional control. The completion of the proposed studies will catalyze our future investigations into elucidating analogous pathways in pathogenic strains, uncovering novel members of these families in bacterial phylogeny, and designing small molecule inhibitors. Project Narrative A number of biochemical processes have promise as antibiotic targets, but efforts towards those goals are hampered in part by a dearth of information regarding their molecular and biochemical details. This proposal will illuminate how bacteria respond and adapt to changes within their environments at a detailed structural and mechanistic level. The findings from these studies will inform future drug discovery effortsto target specific strains of bacteria. | Non-SBIR/STTR | 6project_grants_public |
gen_3dfe84b061865341d3dc5bc536e23ba2 | Investigating Behavioral Mechanisms and Efficacy of a Provider-Directed Intervention for HPV Vaccine Promotion in Real-World Dental Settings | NIH | HEALTHPARTNERS INSTITUTE | 5UG3DE030063-02 | Project Summary Human papillomavirus (HPV) is the leading cause of oropharyngeal cancers in the US. Despite the safety and effectiveness of the HPV vaccine (HPV-V), coverage is far below that for other routine adolescent vaccines and the Healthy People 2030 goal of 80%. HPV-V promotion at dental visits is seen as a prime opportunity to prevent oropharyngeal and other cancers, yet many dental providers are not comfortable doing so due to lack of knowledge and self-efficacy, and fear of harming the patient-provider relationship. Using the NIH Stage Model of Behavioral Intervention Development as our guide, we propose to develop a theory-based intervention to address dental provider barriers to HPV-V promotion, elucidate the intervention's behavioral mechanisms, and test the real-world efficacy of the intervention in catalyzing provider HPV-V promotion. The intervention will consist of 1) provider training about HPV/HPV-V; 2) tailored scripts to aid providers in responding to patient/parent/guardian concerns about HPV-V. During the UG3 phase, we will randomize 21 HealthPartners Dental Group clinics to intervention vs. usual care (UC; n=~131 providers). UG3 aims are to: develop survey measures and pilot-test provider HPV-V promotion training (Aim 1) and tailored scripts (Aim 2); develop measures and methods for monitoring provider fidelity to the training and intervention activities (Aim 3); and draft compliance/study documents and obtain IRB/NIDCR approvals (Aim 4). During the UH3 phase, we will conduct a cluster (clinic)-randomized clinical trial (intervention vs. UC) to test the real-world efficacy of the intervention to increase HPV-V promotion activity (Aim 5). We will assess whether the intervention impacted the three intended behavioral mechanism targets: increased knowledge of HPV/HPV-V; increased self-efficacy for HPV-V promotion; and reduced fear of HPV-V promotion negatively affecting the patient- provider relationship (Aim 6). For each target, we will also assess whether the intervention's effects followed the full mechanistic pathway to the endpoint behavior, HPV-V promotion (Aim 7). Beyond our aims, we will conduct exploratory work examining two additional candidate behavioral mechanisms: adequacy of material resources to support dental providers in promoting HPV-V, and providers' perception that HPV-V promotion comports with their professional identity. We will also conduct an exploratory analysis of the intervention's efficacy in increasing HPV-V uptake (30-day post-visit patient vaccination rates). Our long-term goal is to reduce HPV and oropharyngeal cancer prevalence through HPV-V promotion by dental providers. Significant impact of the project includes: 1) developing the first theory-based behavioral intervention for HPV-V promotion aimed at dental providers; 2) delivering the first evidence of real-world efficacy of such an intervention; 3) illuminating behavioral mechanisms purported to underlie provider behavior change; 4) producing fundamental knowledge to guide future HPV-V promotion intervention development; and 5) advancing the science of behavior change by revealing behavioral principles underlying provider behavior change. Project Narrative Training dental providers to effectively promote the human papillomavirus vaccine (HPV-V) with their patients could prevent future HPV-related cancer of the mouth and throat in those who become vaccinated. This study will develop HPV-V promotion training and tailored scripts to support dental providers in promoting HPV-V. By testing this intervention in a real-world dental setting, this project has the potential to help increase HPV vaccination rates, reduce HPV-related cancers, and serve as a model for other organizations providing dental care. | Non-SBIR/STTR | 6project_grants_public |
gen_bfb6d207a7de183fbd6bd5b55a702c8b | Children's Oncology Group | NIH | UNIVERSITY OF SOUTHERN CALIFORNIA | 3U10CA180899-10S1 | PROJECT SUMMARY Since the introduction of chemotherapy for the treatment of childhood leukemia more than 60 years ago, the prognosis of childhood cancer has improved dramatically. The overall 5-year survival rate for childhood cancers, many of which were uniformly fatal in the pre-chemotherapy era, is now 84%. Progress for a number of childhood cancers, however, has been limited, with approximately 50% of children with acute myelogenous leukemia, 50% of children with high-risk neuroblastoma, and more than 90% of children with brainstem glioma, still succumbing to their disease. In the US, cancer remains the leading cause of death from disease in children greater than one year of age. Moreover, the late effects of cancer treatment, including permanent organ and tissue damage, hormonal and reproductive dysfunction and second cancers, are of special concern, with more than 40% of the 500,000 survivors of childhood cancer (estimated as of 2020) experiencing a significant health related quality of life complication from childhood cancer and its treatment. Thus, despite our advances, development of new therapeutic approaches must be a priority for childhood cancer basic, translational and clinical researchers. The Children’s Oncology Group (COG), the world’s largest organization devoted exclusively to childhood and adolescent cancer research, was founded 20 years ago. The COG’s multidisciplinary research team, comprised of more than 9,000 members, conducts research at more than 220 leading children’s hospitals, universities, and cancer centers. This proposal is for COG, as part of the National Cancer Institute’s (NCI) National Clinical Trials Network (NCTN), to continue its collaborative research work that supports the mission of improving the outcome for all children with cancer. The COG will design and conduct clinical-translational studies for children with cancer that builds on an increasing understanding of the molecular basis for pediatric malignancies and has the highest potential to improve the outcome. Using innovative clinical trial designs suitable for the study of rare diseases, we will study novel therapeutic approaches including but not limited to targeted small molecule drugs, immunotherapies and cellular therapies. The COG research portfolio importantly also includes clinical trials focused on improving the quality of life for children with cancer and childhood cancer survivors. As more than 90% of children diagnosed with cancer in the US are treated at COG member institutions, COG has the ability to offer a diverse population of children with cancer and their families the opportunity to participate in innovative research. This research effort includes allowing for collection and annotation of biospecimens from all children with cancer, providing the foundation for discovery and accelerating the most promising research efforts conducted in laboratories around the world. The proposal is for support of the COG Network Operations Center, scientists who develop and conduct research, and for member sites to participate through enrollment of eligible subjects. PROJECT NARRATIVE The Children’s Oncology Group (COG) is the world’s largest organization devoted exclusively to childhood and adolescent cancer research. Over 220 leading children’s hospitals, universities, and cancer centers across US, Canada and other countries participate in COG research, which is focused on developing better treatments that can improve the cure rate and outcomes for all children with cancer. | Other Research-Related | 6project_grants_public |
gen_3a9e29c825b1e524b6ce58d7286a318f | Dysregulation of Epithelial Metabolism and Regeneration by Sulfite Exposure in Pediatric Ulcerative Colitis | NIH | STANFORD UNIVERSITY | 5K99DK136971-02 | PROJECT SUMMARY The prevalence of ulcerative colitis (UC) in children continues to increase yearly. Recent evidence in pediatric UC patients showed significant mitochondrial impairment in the colon tissues. This is important as optimal mitochondrial activity is required for the solemn function of colonic stem cells that replenish the physical barrier of the colon epithelium. Since patients are constantly exposed to environmental factors such as diet, it is critical to reveal the dietary factors that influence mitochondrial function in the colon epithelium as they would be vital in the management of UC in children. Sulfites are endogenous products of several sulfur-containing compounds, and they are also ubiquitous in our diets as preservatives. My preliminary data in colon organoids derived from pediatric patients showed a detrimental role of sulfite on mitochondrial metabolism and differentiation, with worse metabolic outcomes in samples from pediatric UC patients. My analysis of transcriptomic data from 206 children with UC showed that the Mocs1 gene required for downstream clearance of sulfites in the mitochondria is downregulated in the colon of UC patients, suggesting a potential for inefficient sulfite detoxification in the colon. In this study, I will use patient-derived colon organoids to define how sulfites regulate mitochondrial metabolism and differentiation in health and in UC (Aim 1), reveal the sulfite-induced and sulfite susceptibility chromatin sites in the pediatric colon that explains these metabolic and differentiation anomalies (Aim 2), and how sulfites and the loss of epithelial Mocs1 shape colon biology in the complex gut environment in vivo using physiological relevant models (Aim 3). This award will advance my training in disease models of IBD, epithelial biology, and epigenomics as I work toward establishing an innovative career in regenerative nutrition with a focus on pediatric digestive diseases and continue efforts to enhance diverse representation in the biomedical sciences. PROJECT NARRATIVE Ulcerative colitis continues to increase in children and environmental factors such as diet play a vital role in the management of the disease. Sulfites are potentially harmful compounds that must be detoxified in many cells and tissues, but they are also ubiquitous in our diet as preservatives. To inform future practices on the dietary management of pediatric ulcerative colitis, this study proposes innovative scientific approaches to determine whether sulfites interfere with the normal function of the colon function in health and in UC. | Other Research-Related | 6project_grants_public |
gen_8af4307e8f456cd2f4e522187746d0fc | DMS/NIGMS 1: Multiscale modeling of Notch signaling during long-range lateral inhibition | NIH | CLARKSON UNIVERSITY | 5R01GM152810-02 | The spatiotemporal distribution of morphogens contributes to the organized development of tissues and organs. One model of morphogen distribution is active transport, which includes cell based mechanisms like signaling filopodia. Signaling filopodia facilitate contact between distant cells in order to allow signaling to occur, and support several cell signaling paradigms during development. The proposed project will use multi-scale modeling and biological experiments to test the hypothesis that Notch signaling occurs via filopodia-filopodia mediated cell-cell contacts in vivo. This hypothesis will be tested in three objectives. (1) Investigate the mechanism of Notch activation on filopodia. A mechanical model of distinct modes of filopodia interactions will be used to quantify the forces generated during filopodia mediated signaling to identify the most likely mechanism for Notch activation. (2) Determine how Notch signal is relayed to the cell body. A mathematical model of filopodia in the presence of diffusion and active transport of signals will be developed to quantify the relative importance of each mechanism. We will support our model with genetic approaches and quantitative live imaging. (3) Create a multi-scale vertex model of Notch signaling during bristle cell patterning. We will combine the above molecular and cellular sub_x0002_models of Notch signaling to create a truly multi-scale vertex model of the patterning thorax. This framework will support an in silico, real-time investigation of patterning dynamics via signaling filopodia to identify potential molecular regulators of this process. The success of this proposal will result in a foundational understanding of the mechanisms that drive long-range lateral inhibition during tissue patterning. We will introduce the first multi-scale mechanical model of the fly thorax that allows for cell-driven dynamics of filopodia and real-time activation of Notch. The experimental work proposed here addresses a major gap in our understanding of tissue development and homeostasis: how active cell processes contribute to the distribution and activation of signals. A major challenge for the organized development and homeostasis of tissues and organs is the correct spatial and temporal distribution of signaling molecules. Achieving a correct tissue organization is critical as defects in patterning can lead to human disorders and disease. This project will take both multi-scale mathematical modeling and experimental approaches to investigate how the signaling receptor Notch is distributed and activated by active cell processes during lateral inhibition-mediated tissue patterning events. | Non-SBIR/STTR | 6project_grants_public |
gen_1f1c62ae1c4b6604adeb54867efa0ab2 | In-vivo Assessment of Neuroinflammation in Painful Trigeminal Neuropathy | NIH | MASSACHUSETTS GENERAL HOSPITAL | 1R21DE031410-01A1 | Painful trigeminal neuropathy (PTN) is defined as facial pain in the distribution(s) of one or more branches of the trigeminal nerve, associated with neural damage induced by trauma, viral infection, or other causes. PTN is very unresponsive to medical and surgical treatments. Clearly, a more in-depth knowledge of the molecular pathomechanisms of PTN are in urgent need to improve the management of this disorder. In the last six years our group has demonstrated the presence of increased levels of the 18kDa translocator protein (TSPO, using positron emission tomography) and/or myo-inositol (mIns, using magnetic resonance spectroscopy), in the brains of patients with various chronic pain conditions. Because both TSPO and mIns are overexpressed by glial cells, our results suggest that neuroinflammation might be a pervasive phenomenon that can be observed across multiple, etiologically heterogeneous human pain disorders, but in a disorder-specific spatial distribution within the central nervous system. Despite these advances, the clinical significance of these brain inflammatory signals (e.g., whether neuroinflammation imaging could be used to identify patients more likely to respond to anti-inflammatory therapies) remains to be evaluated. In this exploratory project, we will recruit PTN patients scheduled to receive oral steroid therapy. All participants will be evaluated clinically by an experienced neurologist and, prior to commencing their treatment, receive brain imaging with integrated (i.e., simultaneous) positron emission tomography / magnetic resonance imaging (PET/MRI) and [11C]PBR28, a second-generation radioligand for TSPO, which we have used to demonstrate glial activation in patients with pain or neurodegenerative disorders. After the scan, participants will undergo a 3-week treatment with the steroid prednisone, followed by another clinical/behavioral visit. Clinical characterization will include quantitative sensory testing and questionnaires. Patients’ imaging data will be compared to an existing dataset of healthy controls and chronic pain patients with a different etiology (chronic low back pain) to assess the specificity of our findings to PTN. For Aim 1, we will assess in-vivo neuroinflammation in painful trigeminal neuropathy, using multimodal brain imaging. For Aim 2 we will test the brain neuroinflammatory signals’ ability to predict response to steroid treatment. This work will advance our understanding of the clinical significance of neuroinflammation in chronic pain conditions. While this project is focused on neuropathic pain, identifying the role of glia in the development and maintenance of persistent neuropathic pain and pain-related disability in humans will have important practical implications for the management of a wide range of pain disorders. For instance, it will provide crucial human evidence contributing to rationale for the development of tailored interventions focused on glial modulation. In this exploratory project we seek to extend our investigations of the role of neuroinflammation in human chronic pain to painful trigeminal neuropathy. All participants will receive brain imaging with integrated PET/MRI, before steroid treatment. We will test whether 1) patients demonstrate abnormally elevated PET and MRS markers of neuroinflammation, and whether 2) these brain signals can predict response to steroids. | Non-SBIR/STTR | 6project_grants_public |
gen_89d9b5eca44aed98cbbfd4964eb1f197 | Structure-based inhibition of chemokine signaling in the inflamed pancreas | NIH | MEDICAL COLLEGE OF WISCONSIN | 5R01DK133247-02 | Modified Project Summary/Abstract Section The goal of this project is to establish the role of CCL28 in chronic pancreatitis and use structure-based drug discovery methods to identify small molecule inhibitors of this secreted protein. CCL28 is a mucosal chemokine that promotes tumor growth in a variety of organs by recruiting regulatory T cells (Tregs) that express the G protein-coupled receptor CCR10. Based on our published and preliminary results, we postulate that secretion of the chemokine CCL28 by pancreatic ductal epithelial cells also drives chronic inflammation that progresses to malignant disease. We hypothesize that inhibition of CCL28 activity will alter the pancreatic mucosal microenvironment in a manner that reduces pre-malignant inflammation and enhances the activity of existing chemotherapeutics and immunotherapies. To achieve this objective, we propose three conceptually linked but experimentally independent specific aims. First, we will test our mechanistic hypothesis in animal models of chronic pancreatitis, to demonstrate experimentally that CCL28 activity through CCR10 plays a key role in the fibroinflammatory response in vivo and develop an in vitro model that can be used to screen promising inhibitors (aim 1). Key elements of CCL28 recognition by its G protein-coupled receptor CCR10 will be mapped in detail using NMR and molecular modeling, and we will define the complete intracellular signaling profile of this chemokine receptor using state-of-the-art assay platforms for receptor pharmacology (aim 2). Using the solved NMR structure of CCL28 and knowledge of its sulfotyrosine binding pocket, we will employ a structure-based strategy developed in the Volkman lab that enables the discovery of small molecules that bind a specific chemokine target and inhibit cell migration (aim 3). Collectively, the proposed studies will provide fundamental advances in our understanding of (1) Treg function and pathophysiology in the pancreas, (2) the structural basis for ligand-receptor selectivity and GPCR pharmacology in a largely unexamined chemokine signaling axis, and (3) the druggability of a mucosal chemokine at the tumor-promoting interface of chronic inflammation and neoplasia. PROJECT NARRATIVE The research proposed in this application is relevant to public health because chronic pancreatitis is a progressive, incurable inflammation that leads to permanent organ damage and cancer. The proposed work will define key roles for pancreatic immune messengers that control damaging inflammatory responses and use structure-based drug discovery approaches to disrupt disease progression and improve patient health. | Non-SBIR/STTR | 6project_grants_public |
gen_b92f76df87a8968dbe6ca1238c2b1781 | Age Differences and Mechanisms of Ketogenic Diet Induced Bone Loss | NIH | UNIVERSITY OF CALIFORNIA AT DAVIS | 5K99AR081401-02 | Project Summary Ketogenic diet (KD), a high fat low carbohydrate diet is used to treat intractable epilepsy, is becoming increasingly popular for weight management, and it can potentially slow cognitive ageing and alleviate symptoms of neurological disorders such as stroke, Parkinsons disease, and Alzheimers. However, KD also causes bone loss and increases fracture risk in children. It is not known if KD causes bone loss in adults. Based on prior studies, it is also possible that KD may reduce the ability for exercise to increase bone strength. The mechanisms responsible for KD bone loss have not been identified. Determining if β-hydroxybutyrate (BHB), the most abundant ketone body is linked to bone loss is important, because this molecule is thought to lay a large role in the neurological benefits of KD. This project will use a mouse model to evaluate age differences in ketogenic diet induced bone loss, determine if KD decreases the ability of exercise to make bone stronger, and investigate if BHB causes bone loss. Aim 1 will determine how age and diet duration affect the magnitude of KD induced bone loss and decrease in bone strength. Aim 2 will evaluate if KD reduces the ability of exercise to increase bone strength and if this is mediated by muscle and tendon. Aim 3 will focus specifically on defining the role of BHB in bone loss. In the long term, this project will help clarify how KD affects bone, and it can contribute to the use of KD or BHB supplementation to deliver neurological benefits without increasing fracture risk. As a clinical researcher, I strive to develop therapies to improve skeletal health, and as a biological anthropologist, I use skeletal remains to reconstruct the behavior and health of past people. Through the K99/R00 career development award, I seek to combine the anthropological and biomedical strands of my research career by examining the combined effect of ketogenic diet and exercise on bone health throughout life. During the fellowship, I will receive training in molecular biology and laboratory skills essential for the study of cellular responses to diet and exercise. I will also expand my knowledge of muscle and tendon, gaining the ability to conduct innovative interdisciplinary research that achieves new perspectives on how exercise and diet affect bone strength. UC Davis is an unparalleled location for conducting the proposed project and training. I will have access to cutting edge facilities and equipment. Through numerous seminars, workshops, and training opportunities I will interact with faculty, students, and staff, broadening my understanding of skeletal health. Through the K99/ R00 I will develop an innovative interdisciplinary research career that explores the relationship between diet, behavior, and health in past human populations and contributes to the development of therapies that use diet and exercise to decrease fracture risk. Project Narrative Ketogenic diet is popular for weight management and may help treat neurological disorders, but it may also cause severe bone loss that increases fracture risk. This project contributes to public health by characterizing the effect of ketogenic diet on bone strength throughout adulthood, if KD prevents exercise from increasing bone strength, and investigating if ketone bodies are a primary mechanism driving bone loss. In the long term, the findings can improve understanding of how ketogenic diet contributes to fracture risk and the development of therapies to prevent bone loss on a ketogenic diet. | Other Research-Related | 6project_grants_public |
gen_dbf664ecc4d9500fef1cb70ed986bc03 | Access for All in ALS (ALL ALS) West Clinical Coordinating Center | NIH | ST. JOSEPH'S HOSPITAL AND MEDICAL CENTER | 3OT2NS136939-01S1 | Abstract Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, ultimately fatal, neurodegenerative disease with approximately 32,000 cases in the United States (US). With an estimated lifetime risk of 1/400, an average of 5,000 new US cases are diagnosed every year. A public private partnership (PPP) can be leveraged to rapidly advance regulatory science and scientific research to support and accelerate the development of effective, new drugs to extend life for people living with ALS. Given the numerous failures in translating laboratory results into clinically effective therapies, the generation of high quality, diverse and longitudinal patient-derived biosamples and resulting downstream data is critical for our success. To this aim, our proposed ALS Clinical Research Coordinating Center (ALSynCC) builds and expands upon the ongoing success of our Target ALS Natural History study and consortium and will include five sample/data repositories linked to detailed clinical information: 1) a biofluid repository that contains cross-sectional and longitudinal biofluid sample from asymptomatic and symptomatic patients where samples are linked to detailed clinical information, whole genome sequencing, at-home measures of speech and respiratory function, 2) a longitudinal study of asymptomatic mutation carriers called the PREVENT study that will be expanded to include participants across the country, 3) an MRI- based neuroimaging repository to be newly implemented, 4) at-home measures of speech and respiratory function, and 5) a large, well-annotated post-mortem tissue biobank containing brain, spinal cord and muscle tissue from patients. Each of these repositories is led by highly trained physicians and scientists, experts in the field. This use of multi-modal and multi-compartment sampling from each patient is a powerful method to characterize disease processes and will improve translation of diagnostics, therapies, and outcome measures to the clinic. These highly curated biosamples and data resources are de-identified, linked to detailed clinical information, and will be made immediately available to the research community through the Consortium data portal. Our Target ALS natural history program is a successful public-private partnership between academic and industry leaders and closely mirrors the overall goals of the proposed ALS Clinical Research Consortium and functions of the Coordinating Center (CC). A unique aspect of our model is to collect data and samples from asymptomatic individuals at risk for ALS, as well as from symptom onset to autopsy to provide a comprehensive sample and dataset for future research studies. We will utilize our knowledge and tremendous success to expand on this existing infrastructure to maximize ALS patient enrollment in a highly synchronized research platform called ALSync that is administered by the ALSynCC. This nationwide ALSync network will work closely with NIH and other partners to coordinate and bridge ALS research and clinical care across the country to: a) ensure inclusion of male and female patients of all racial and ethnic backgrounds, b) to improve access to clinical research and trials, c) to generate high quality patient samples and data, and d) to efficiently share data and analysis tools to accelerate research in the broader ALS community. | Other | 6project_grants_public |
gen_fb19479292a3fec4f3815e79ef477573 | Maximizing tumor responses to targeted radiotherapy with a conditionally activated membrane binding probe | NIH | UNIVERSITY OF CALIFORNIA, SAN FRANCISCO | 1R01CA279328-01A1 | Project Abstract: The recent FDA approvals (Lutathera, Azedra, Pluvicto) and the swell of promising experimental agents in clinical trials underscore the surging enthusiasm to investigate molecularly targeted radiotherapy (TRT) as a treatment modality for cancers. However, tumor responses to TRTs are often transient and/or variable among patients. Thus, there is an urgent unmet need to develop new strategies to maximize the therapeutic benefit of TRT for cancer patients. For the past several years, the nuclear medicine field has prioritized developing low MW small molecule or peptide radioligands (RLTs) that rapidly exit the bloodstream to minimize host toxicity. However, tumoral responses to RLTs are limited by several factors, including heterogeneous target expression among tumors, dissociation or degradation of ligand/receptor complexes, and incomplete target saturation due to low mass doses and infrequent repeat dosing. Thus, exploring new strategies beyond RLTs for the tumoral delivery of radioisotopes is a worthwhile goal. We have approached this challenge by developing a new class of radiopharmaceuticals termed “restricted interaction peptides” (RIPs) which are linear and unstructured low MW peptides that are internally cleaved by a tumor endoprotease of interest to unmask a radiolabeled, helical membrane binding peptide. Once liberated, the radiolabeled helical peptide immediately and irreversibly attaches to a nearby phospholipid membrane in the tumor. Using PET, we have found that RIPs may have several properties advantageous for TRT, including catalytic amplification of tumor uptake and long persistence of the radioisotope in tumors due to the stability of the peptide/lipid membrane interaction. Thus, RIPs offer an unusual combination of the desirable safety profile characteristic of a low MW RLT with a high tumoral uptake more typical of a large MW TRT. Collectively, these findings provide a strong scientific rationale to test for the first time if radiolabeled RIPs can be effectively leveraged to treat tumors. Over three specific aims, we will evaluate the antitumor effects of a novel RIP termed “FRIP2” by coupling it to a representative β- (Lu-177) or alpha (Ac-225) emitter. Furthermore, we will translate 64Cu-FRIP2 into patients to test the safety, dosimetry, and pharmacokinetics of the platform while also evaluating the feasibility of tumor targeting. In summary, this project represents the first use of a conditionally activated membrane binding probe for TRT, which may overcome the well documented shortcomings of conventional RLT. Project Narrative: The overall goal of this project is to develop a new platform for the precision delivery of targeted radiotherapies to tumors. Over three specific aims, we will evaluate if proteolytically activated membrane binding peptides can be leveraged to deliver a more substantial dose of ionizing radiation to tumors than conventional radioligand therapies. In parallel, a lead drug candidate conjugated to copper-64 will be translated into cancer patients to assess safety, dosimetry, and pharmacokinetics, as well as the feasibility of tumor targeting using PET. | Non-SBIR/STTR | 6project_grants_public |
gen_07bf905f04d124a3053d26734d6e9569 | Investigating mechanisms of oxidized phospholipid-mediated dysregulation of regulatory T cells in atherosclerosis | NIH | VANDERBILT UNIVERSITY MEDICAL CENTER | 1R21AI176219-01 | SUMMARY The important role of adaptive immunity in cardiovascular disease (CVD) is established. However, much is still unknown regarding how adaptive immunity can be manipulated to benefit CVD outcomes. Atherosclerosis, the most common form of CVD, is characterized by accumulation of lipid and immune cells in the artery wall that leads to plaque formation. Recent single cell RNAseq data in human atherosclerotic confirm that T cells make up a large fraction of the cell population in lesions. T cells are important to the atherosclerotic process as effector T cells (Th1 and Th17) seem to promote plaque growth and instability whereas regulatory T cells (Tregs) are critical for inhibition of atherogenesis and induce regression of established plaques. In humans, Tregs are associated with plaque stability and protection against cardiac events. These data are consistent with mouse models that show Tregs numbers in plaques decrease as atherosclerosis progresses and remaining Tregs Th1-like inflammatory phenotype. Why Tregs dedifferentiate or become dysfunctional in atherosclerosis is not completely understood, but recent studies suggest that oxidized low density lipoprotein (oxLDL) may play a functional role in this process. Preliminary studies from our laboratory examined whether oxidized phospholipid (oxPAPC), associated with atherosclerosis and oxLDL, could affect Treg differentiation, stability, and function. Our preliminary data show that, oxPAPC reduced Treg viability and increased expression of the Th1-associated transcription factor T-bet and production of IFN- thus promoting the Th1-like Treg phenotype. This effect was Treg-specific as cells skewed in Th1 or Th17 conditions were not affected by oxPAPC. These oxPAPC Tregs were also less suppressive in vitro. The effect of oxPAPC on Tregs was partially dependent on CD36 and IFN-. Interestingly, IFN- has been shown to destabilize Tregs in the tumor microenvironment and data in this proposal show that neutralization of IFN- in Treg skewing cultures inhibits expression of T-bet in FoxP3+ cells. Therefore, we hypothesize that oxidized phospholipids associated with atherosclerosis induce an effector- phenotype in Tregs that is partially dependent on IFN-. To test this hypothesis we propose two specific aims. The first will examine the role of CD36/TLR and IFN- signaling on the dysregulation of Tregs in the presence of oxPLs and determine if these Tregs can protect from atherosclerosis in vivo. The second, will examine whether CD36 or responses to IFN- are responsible for destabilization of the Treg compartment during atherosclerosis in vivo. This work will uncover novel information on the role of oxidized phospholipid on Treg dysfunction in atherosclerosis and will provide valuable insight for future clinical interventions. NARRATIVE Regulatory T cells have been shown to play important protective roles in atherosclerosis, including being critical for plaque regression. It is known that the atherosclerotic environment dysregulates regulatory T cells making them more inflammatory. This proposal hypothesizes that oxidized phospholipids that accumulate in plaques are mechanistically important for the dysregulation of regulatory T cells in atherosclerosis. | Non-SBIR/STTR | 6project_grants_public |
gen_4aea3d01a372c2fa2d3a1b2fde3c8f06 | VRC Inhibiting p38 to Prevent and Restore Corneal Scarring | NIH | SCHEPENS EYE RESEARCH INSTITUTE | 1R01EY035947-01 | Project Summary Research Idea/Rationale: Corneal scarring (or fibrosis) from corneal puncture injury or laceration is one of the leading etiologies for blindness. Currently, none of the potential treatments for corneal fibrosis have been FDA- approved and none have been shown to reverse an existing corneal scar. Therefore, finding a therapeutic with the ability to reduce or control scarring, but that is not invasive or has minimal adverse effects, remains an unmet need. Studies from both in vivo and in vitro models have demonstrated that transforming growth factor- beta1 (TGF-beta1) is one of the key factors that drive corneal scarring after incision injury. However, TGF- beta1 is not an ideal therapeutic target for fibrotic diseases due to its importance in normal physiological processes (e.g., proliferation and migration). Objective/Hypothesis: During our preliminary in vitro studies, we found that an inhibitor for the TGF- beta/p38MAPK (p38)-signaling pathway, SB202190, almost completely blocked human corneal scar formation. This discovery led us to examine if p38 inhibitor could prevent the transformation of human corneal fibroblasts (normal active stromal cells) into myofibroblasts, a stromal cell phenotype that is involved in scarring. We found that p38 inhibitor accelerated the conversion of these myofibroblasts back to their normal phenotype. We hypothesize that the p38 inhibitor, SB202190, can be used as a local therapeutic for preventing and reversing corneal scarring. Specific AIMS: Aim 1:Safety evaluation of SB202190. AIM 2: Determine the therapeutic effect of the SB202190 to prevent corneal scar formation. AIM 3:Determine if an established corneal scar can be reversed by treatment with SB202190. Study Design: In this model, a 3mm perforating incision injury will be made in the center of the right cornea of Sprague-Dawley rats. For AIM 1, treatment will be applied immediately after wound creation and reevaluated at 24 hours to ensure SB202190 does not cause wound healing delays. Cytotoxicity will also be evaluated in human corneal fibroblasts. For AIM 2, SB202190 or control PBS will be applied locally for 1-15 days as follows: eye drops (topical) will be applied 3 times daily or subconjunctival injection once every three days. For AIM 3, the injured corneas will be allowed to heal for 14 days. Rats will be randomly grouped and treated locally as in AIM 1 for 0-8 weeks. Corneas will be examined for haze and proteins associated with scarring at 0-14 days post-treatment in AIM 2 and 0-8 weeks post-treatment in AIM 3 using the following techniques: 1) Slit lamp and OCT examination; 2) TEM and standard Hematoxylin and Eosin stain 3) Indirect- immunofluorescence and Western blot. Absence, or at least significant decrease, of haze and scar proteins will indicate that the inhibitor prevented scarring in the animal model. The purpose of this research is to determine if a mitogen activated kinase (MAPK) inhibitor can prevent or reverse corneal scarring. There are currently no treatments to reverse a corneal scar, which results in a permanent loss of vision. | Non-SBIR/STTR | 6project_grants_public |
gen_dbf5f7e645b28dfef713c5e7cfd12c5b | DiversitySupp-ONES-Prenatal Phthalate Exposure | NIH | SEATTLE CHILDREN'S HOSPITAL | 3R01ES033785-01A1S1 | Project Summary Spontaneous preterm birth (sPTB) comprises the majority of preterm births (60%) and is a leading case of newborn morbidity and death and a predictor of adverse health outcomes. Despite its high prevalence, there is a limited understanding of how the in-utero environment contributes to the etiology of sPTB. Phthalates are ubiquitous endocrine disrupting chemicals that induce gene expression and physiological changes within the placenta. Epidemiological studies identify a consistent positive relationship between prenatal phthalate exposure and preterm birth. The goal of this study is to develop placental molecular signatures that can be used to mechanistically link prenatal phthalate exposure and sPTB. Placental molecular signatures can explain functional differences related to sPTB and identify targets for clinical and therapeutic interventions, including modifiable risk factors such as environmental exposures. Our research team has generated the largest placental transcriptomics dataset to date (N=760 samples) and has used this to develop transcriptomic signatures of prenatal phthalate exposure and sPTB. This study will expand our existing transcriptomic signatures to include microRNAs, which are essential to a complete molecular signature because they are highly stable, have been linked to a number of environmental exposures, and are secreted into maternal circulation where they may serve as biomarkers. Candidate microRNA studies have identified correlations between prenatal phthalate exposure and expression of placental microRNAs, but a comprehensive assessment is needed to fully understand the role of placental microRNAs in phthalate mediated toxicity. Moreover, despite the potential importance of placental microRNAs as a biomarker of sPTB, there has not been a comprehensive analysis. In this proposal, we seek to fill these research gaps and apply innovative computational biology strategies with rigorous epidemiological approaches to gain insight into the mechanistic links between prenatal phthalate exposure, placental function, and sPTB. In aim 1, we will generate microRNA data on placental samples and use this to generate a signature of prenatal phthalate exposure. We will use the matched microRNA-mRNA sequencing data to construct a global placental microRNA-mRNA network, which we will apply to identify connections between microRNAs and genes whose placneta expression is associated with different phthalate metabolites. In Aim two, we will develop a multi-omic molecular signature of sPTB using our placental microRNA-mRNA network. In aim 3, we will examine the role of the placenta as a mechanistic link between prenatal phthalate exposure and sPTB by interdisciplinary strategies including an integrated pathway analysis and a formal mediation analysis. Findings from this study will inform chemical toxicological risk assessment and policy to reduce health impacts due to phthalate exposure in pregnancy. microRNA signatures of sPTB may serve as functional biomarkers of sPTB since they can be secreted into maternal circulation and be targets for clinical and therapeutic intervention in the future. PROJECT NARRATIVE Spontaneous preterm birth is a highly significant but understudied perinatal outcome that comprises the majority of premature births and has been linked to prenatal exposure to environmental chemicals, including phthalates. This study will investigate the mechanistic link between prenatal phthalate exposure, placental dysfunction, and spontaneous preterm birth using multi-omics data generated in the placenta. Findings from this study will inform chemical toxicological risk assessment and policy, as well as aid in the development of potential biomarkers of spontaneous preterm birth. | Non-SBIR/STTR | 6project_grants_public |
gen_eaebaaa965bb7b7dfb2051aab516faf3 | Dissecting the complex role of microglia states in glaucoma | NIH | UNIVERSITY OF ROCHESTER | 1R01EY035093-01 | Glaucoma is a very common age-related neurodegenerative disease characterized by the death of the retinal ganglion cells. Despite its prevalence, there are no neuroprotective treatments for glaucoma. The only current treatment is lowering intraocular pressure which unfortunately does not prevent or restore vision loss in many patients. Due to extensive research in both glaucoma patients and animal models of glaucoma, much is known about glaucomatous neurodegeneration, both at the physiological and molecular levels. However, despite this, we still lack a molecular understanding of the early pathological events that injure RGCs as a result of ocular hypertension. Microglia cells are a major component of the neuroinflammatory response in neurodegenerative diseases and after injury to the central nervous system. In fact, microglia cell response is thought to play key roles in many neurodegenerative diseases, including, Alzheimer’s disease, Parkinson’s disease, Huntingtin disease, and Amyotrophic Lateral Sclerosis. Work in other systems has shown that microglia can act be both protective and detrimental in the disease process, and possibly these two actions could take place sequentially in the same disease. Recent studies using modern sequencing technology has shown that microglia exist in different molecular states which correspond to their role in disease. A major, well-supported hypothesis in glaucoma research is that microglial cells are critical for maintaining retinal ganglion cell viability after a glaucomatous insult. However, the importance of microglia in an ocular hypertensive model of glaucoma has not been critically tested. In this application we propose to test the hypothesis that different activated states of microglia play distinct roles in glaucoma dependent upon stage of disease. Specifically, in two ocular hypertensive glaucoma mouse models, we will: (1) Determine if the role of microglia activation varies with disease stage, (2) define and test the importance of different molecular states, and (3) determine the role of microglial derived neurotoxic cytokines. Overall, this proposal we will define novel mechanisms by which microglia states modulate glaucoma onset and progression leading to novel candidates for therapeutic evaluation. Microglial cells, which are key components of the neuroinflammatory response to neuronal injury, are thought to be important mediators of retinal ganglion cell viability in glaucoma. In this application, we will critically test the importance microglia in glaucoma; furthermore, we will use advanced RNA sequencing technology to determine if there are different states of microglia—ones that are potentially beneficial or detrimental to retinal ganglion cell survival. Together, these data will determine if targeting microglial activation is a potential treatment for glaucoma. | Non-SBIR/STTR | 6project_grants_public |
gen_626d93cbe2761a044fb9275b928dea42 | The role of adaptive immunity in organophosphate induced CNS injury | NIH | ROSKAMP INSTITUTE, INC. | 5R21NS131162-02 | Chlorpyrifos (CPF) is an organophosphate (OP) pesticide, classified as a chemical threat agent by the Department of Homeland Security because it can cause neurotoxicity if released into the civilian populations. Under these circumstances, CPF and similar OP chemicals have a potential to cause long-term chronic multi symptom illnesses (CMI), such as myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). To date, many victims of the Tokyo subway sarin gas attack are still experiencing chronic health problems consisting of cognitive difficulties, headaches, muscle weaknesses and fatigue, which negatively impact their quality of life. Such symptoms are reported by individuals exposed to OP pesticides and are thought to be caused by maladaptive immune responses7. As such, this proposal will investigate the role of CPF in maladaptive immune responses to develop future approaches that mitigate long-term morbidity associated with CMI. Ordinarily, a small chemical is not antigenic, but when it forms adducts with endogenous proteins, it can be recognized by the immune system as a foreign threat agent and provoke an adaptive immune response. Our prior work in this area shows that certain pesticide metabolites can form adducts with proteins and then elicit an adaptive immune response, activating T- and B-cells that ultimately contribute to the production of antibodies against them and corresponding with brain inflammation. Accounts of acute CPF poisoning in humans support this notion by showing that CPF or CPF-oxon (CPO) can form adducts on several amino acid residues in human albumin, which are considered biomarkers of OP exposure. However, it is unknown whether these CPF/CPO-protein adducts have a role in activating the immune system. We therefore hypothesize that CPF/CPO-protein adducts formed in vivo after CPF exposure can activate T-cell and B-cell responses, resulting in antibody production. We propose that CPF-protein specific antibodies may cross-react with brain proteins and contribute to the development of chronic autoimmune disorders. The proposed work builds upon an existing scientific premise of pesticide-mediated maladaptive immune responses. These studies will characterize whether acute CPF administration stimulates immune cells and whether this corresponds with activation of the microglia and astroglia and neuroinflammation after CPF exposure. We will determine whether brain immune activation is associated with formation of CPF/CPO-protein adducts. We will examine the presence of immune cells that recognize CPF/CPO-modified proteins and antibodies against them to determine if blood antibodies can cross-react with brain proteins. Understanding the mechanisms of OP- induced CMI will facilitate the development of countermeasure efforts that target the immune system in order to minimize long-term morbidity associated with such illnesses in civilian populations following a mass chemical attack with CPF or similar chemicals. Organophosphate (OP) pesticides, such as chlorpyrifos (CPF) are among the most toxic chemicals known to mankind and their use in Syria by terrorist on innocent civilians highlights the need to identify countermeasure approaches against these chemical threat agents. Many of the symptoms experienced by civilians intoxicated with OP chemicals resemble those associated with autoimmune disorders and, therefore, we will identify mechanisms associated with OP induced immune dysfunction. The proposed work will help identify mechanisms which could lead to future development of treatments that can improve the health of individuals who may suffer from chronic multisymptom illnesses after being exposed to such chemicals. | Non-SBIR/STTR | 6project_grants_public |
gen_57739f81773cb9e1b31126260788d861 | Dysregulated neutrophil subpopulations as a driving mechanism of liver and gastrointestinal disease in HIV-1-infected individuals | NIH | UNIVERSITY OF ALABAMA AT BIRMINGHAM | 5R01DK135413-02 | 7. PROJECT SUMMARY / ABSTRACT. Gastrointestinal (GI) mucosal damage and destruction of the gut epithelial barrier are the defining features of the pathogenesis of HIV-1 infection. Accumulated evidence indicates that neutrophils play a critical role in the gastrointestinal and liver damage in HIV-1 infection. Neutrophils infiltrate the GI tract in HIV-1- infected individuals at high levels and their presence is associated with damage to the epithelial barrier, elevated epithelial permeability, and increased disease severity in animal models and HIV-1-infected patients. In this application, we propose that microbial translocation and the resulting systemic innate immune dysregulation mediated by changes in neutrophil subpopulations in circulation, gut-associated lymphoid tissue (GALT), and liver plays a fundamental role in HIV-1 disease progression. The overall objectives of this proposal are to define the role of neutrophil subpopulations and NETosis as driving mechanisms of gastrointestinal and liver damage in HIV-1 infection and to identify the mechanisms responsible for chronic neutrophilic activation in HIV-1 infection in order to reveal the specific checkpoints for intervention. Our central hypothesis is that HIV-1 infection is associated with the induction and expansion of specific neutrophilic subpopulations with increased capacity to produce reactive oxygen species (ROS) and undergo NETosis. ROS and NETs released from activated neutrophils promote damage in the GI mucosa and liver and drive the progression of HIV-1 infection. This hypothesis has been formulated on the basis of our preliminary data and recently published reports demonstrating the critical role of neutrophils in HIV-1 infection. In preliminary studies, we optimized methods for detailed neutrophil characterization and demonstrated that neutrophils from HIV-1-infected individuals display an activated phenotype, immunosuppressive properties, specific transcriptional profile, increased rate of degranulation, and a high capacity to undergo NETosis. Specific properties of the newly identified neutrophil subpopulations strongly indicate that they play a critical role in damaging GI mucosa and the pathogenesis of liver disease in HIV-1-infected individuals. We propose to determine the effect of induction of specific neutrophil subpopulations on the progression of liver disease in ART-treated HIV-1-infected individuals, to identify specific properties of neutrophil subpopulations in the GALT and liver of HIV-1-infected individuals, and determine whether the innate immune dysregulation in these tissues is associated with a shift in the ratio of tissue macrophages exhibiting M1 versus M2 phenotype resulting in lowered efferocytosis and accumulation of neutrophils undergoing NETosis. The significance of the proposed studies is that once the role of neutrophils in the progression of HIV-1 infection is defined, neutrophil activation and induction of pathogenic populations can be pharmacologically targeted. 8. NARRATIVE In this application, we propose to determine the effect of the newly identified neutrophil subpopulations on the progression of liver and gastrointestinal disease in ART-treated HIV-1-infected individuals. Since functional dysregulation of the neutrophil population and its impact on the innate immune balance can be pharmacologically targeted, understanding the underlying pathogenic mechanisms will lead to novel treatment approaches in HIV-1 infection and other chronic inflammatory conditions. | Non-SBIR/STTR | 6project_grants_public |
gen_fa5dd5828e01036e27bb4a9acd215fa4 | Monoclonal Antibody to Combat Pseudomonas Aeruginosa | NIH | EMORY UNIVERSITY | 5R01AI176545-02 | PROJECT SUMMARY/ ABSTRACT. Pseudomonas aeruginosa is a Gram-negative, opportunistic pathogen that infects immunocompromised individuals, especially in the hospital setting. This bacterium is an important pathogen in people with weakened immune systems, injuries, and other underlying physiologic dysfunctions. P. aeruginosa is responsible for up to 20% of all hospital-acquired pneumonias. It is one of major causes of nosocomial infections and has been noted to be one of the most common bacteria co-infecting patients with COVID-19 or causing super-infections following COVID-19 infections. Despite improvements in antimicrobial therapy and hospital care, P. aeruginosa bacteremia and pneumonia remains fatal in about 30% of cases. P. aeruginosa is also the leading cause of chronic life-threatening lung infections in cystic fibrosis patients. This bacterium is naturally antibiotic resistant and infections are notoriously difficult to treat once established, with no vaccine available. We propose using the abundant and essential protein, elongation factor-Tu (EF-Tu), as an antibody target for P. aeruginosa. While best known for its role in protein synthesis, work from our group and others indicate that EF- Tu can be surface-exposed on P. aeruginosa. Our PRELIMINARY RESULTS show that P. aeruginosa EF-Tu is immunogenic in mice and protective in a murine model of acute P. aeruginosa pneumonia. We have generated a mouse monoclonal antibody to EF-Tu promotes partial clearance of P. aeruginosa in this model. This “Partnerships for the Development of Novel Therapeutics to Combat Select Antibiotic Resistant Bacteria and Fungi” (RFA-AI-22-028) and represents a collaborative effort between Dr. Joanna Goldberg at Emory University in Atlanta, GA (the PI), Dr. Vu Truong at Aridis Pharmaceuticals in Los Gatos, CA (industrial partner), Dr. Marion Pepper from the University of Washington, Seattle, WA and Dr. Sebastian Alberti from the University of Balearic Islands in Palma, Spain. Using spleens from mice we will immunize mice with EF-Tu using protocols optimized for protection and blood from humans that have high titers of antibodies to EF-Tu, we will obtain B- cells that will be screened at single cell level using flow cytometry and a nanoculture microfluidic array, respectively, to identify monoclonal antibodies specific for P. aeruginosa EF-Tu, followed by functional screens for binding and phagocytic killing of P. aeruginosa. We will also test these monoclonal antibodies for efficacy in mouse models of infection as an initial step toward future clinical studies. We believe the studies proposed here represent the appropriate first steps towards developing a new passive reagent that could be given to P. aeruginosa-infected patients regardless of the nature of the infecting strain and associated antibiotic-resistance profile, as well as the immune status of the patient. By the completion of this project, we will have generated and validated 10 murine-derived and 10 human-derived monoclonal antibodies to EF-Tu. Partnering with Aridis Pharmaceuticals for this project will keep us focused on our goal of developing these reagents for human use. PROJECT NARRATIVE. New approaches to fight Pseudomonas aeruginosa infections are desperately needed. We have already generated a monoclonal antibody to the conserved essential protein EF-Tu and shown it to be able to decrease colonization in a murine model of acute P. aeruginosa pneumonia. Here we will screen spleen cells from immunized mice and blood from humans with high titer antibodies to EF-Tu at single cell level using flow cytometry and an innovative nanoculture microfluidic array, respectively, to generate additional monoclonal antibodies to EF-Tu with even more efficacy based on in vitro binding assays, opsonophagocytic killing, and mouse challenge studies, as a first step to develop novel reagents to treat infections caused by this opportunistic pathogen. | Non-SBIR/STTR | 6project_grants_public |
gen_b262bb005cfb9841e5694f8f8724fc7d | Outreach Core | NIH | UNIVERSITY OF CALIFORNIA, SAN DIEGO | 1U54CA285115-01 | PROJECT SUMMARY/ ABSTRACT – OUTREACH CORE The Cancer Research and Education to Advance HealTh Equity (CREATE) Partnership’s geographic area is composed of San Diego and Imperial Counties, the two U.S.-Mexico border counties in California which include ~3.5 million residents. This region is a majority-minority area, with extensive cultural, linguistic, and racial/ethnic diversity. Disparities in cancer risk factors, screening, treatment and survivorship continue to persist nationally and within the Partnership’s region, particularly among underserved groups. Generally, medically underserved individuals are diagnosed with cancer at a later stage and have poorer survival compared to individuals who are more affluent. Reducing these cancer disparities requires capacity development for evidence-based practice and community-engaged research and practice dissemination. There is also a need for greater engagement and training of researchers from under-represented backgrounds to collaborate with community organizations that serve this population. Further, communities rarely have opportunities to develop research programs and implement evidence-based practices in collaboration with expert clinicians and researchers. Using innovative approaches that are integrated into all Partnership components, the Outreach Core addresses these gaps through its overarching goal: To integrate community engagement principles into all CREATE Partnership activities. To accomplish this, the Outreach Core will adopt the Community-to-Bench model, which provides an intentional effort to ensure community engagement and bi-directional dialogue. The following Specific Aims are proposed: 1) Advance cancer health equity by facilitating and supporting catchment area-relevant research and community-engaged research including but not limited to Partnership-sponsored research projects; 2) Provide support to community health centers for screening, early detection, linkage to care and referral pathways; 3) Foster community engagement by conducting needs assessments, disseminating evidence-based practices and resources and implementing educational and outreach activities; and 4) Contribute to the Partnership’s educational mission by providing educational opportunities to undergraduate, graduate and medical students, as well as early-stage investigators in areas of community-engaged research and cancer disparities. Working closely with the Community Advisory Board, the OC will build upon existing community partnerships to further expand the Partnership’s outreach activities with underserved communities and to support research in catchment area priority cancers. To conduct this work, the Core has assembled a multidisciplinary team with expertise in cancer disparities research, prevention, multilevel interventions, and community health system interventions. The Core’s collaborative approach is responsive to community needs and will improve cancer health equity in the Partnership’s region and beyond. | Research Centers | 6project_grants_public |
gen_ff073d6a300e3428df30e17b0cccc000 | Characterizing Vision Impairment and Its Impact on Independence in Older Adults | NIH | UNIV OF NORTH CAROLINA CHAPEL HILL | 5R01EY033907-02 | ABSTRACT Vision impairment (VI) is a significant public health problem among older adults. VI impacts a broad range of activities and is associated with reduced quality of life and poorer physical and mental health. As the population is aging, VI will become an increasingly important problem. Yet, data on VI in the oldest old (aged 80+) are limited. Available data suggest that a significant proportion of older adults have uncorrected refractive error (URE) – a condition that is easily addressed with spectacles – as well as vision-limiting cataract. While distance visual acuity is the most commonly measured aspect of VI, contrast sensitivity (CS; the ability to differentiate objects in low light settings) is also important for many activities. However, few population-based studies have measured contrast sensitivity in older adults; data are needed to understand the impact of distance acuity and contrast sensitivity deficits in the oldest old. In this study, we will add distance visual acuity and CS tests to an already funded, one-time visit in the homes of 4,200 older women who have participated in the Women’s Health Initiative (WHI) for nearly 30 years. These data will be used to characterize the prevalence of VI in the oldest old (Aim 1) and to assess the impact of VI on social, psychosocial, cognitive, and physical health indicators (Aim 2). Given the unique opportunity to link to an ongoing cohort, we will be able to describe cross-sectional associations (Aim 2a) and to prospectively evaluate trajectories of functional decline (Aim 2b). Additionally, we will utilize the WHI-Medicare claims link to analyze associations between VI and healthcare utilization and costs (Aim 2c). We will recommend eye care for individuals with VI and will characterize differences between those who seek care vs. those who do not and will examine reported barriers to care (Aim 3a). By engaging a patient care navigator, we aim to help facilitate care for those who do not initially seek it, and we will evaluate the potential impact of patient care navigation on improving uptake of needed eye care (Aim 3b). Adding these vision measures to the rich repository of WHI data provides a unique opportunity to evaluate the role of VI on a broad range of health domains. Project results will be used to develop interventions to improve vision health in older adults, with the ultimate goal of extending independence and improving quality of life in older adults. Renowned vision experts have highlighted the need for obtaining quality estimates of VI in the oldest old, in order for health systems to prepare for serving the growing population of affected individuals. This innovative project fills an important gap in our knowledge of VI and its impact on the lives of our society’s oldest members. PROJECT NARRATIVE Older adults are the fastest growing segment of the population. While vision loss is common in this age group, information on the true prevalence of vision impairment (distance visual acuity impairment and contrast sensitivity deficits) and the impacts that these impairments have on daily function and quality of life is limited. The goal of this project is to determine how common these impairments are in women aged 80+ and the impact that they have on physical and mental well-being. | Non-SBIR/STTR | 6project_grants_public |
gen_a66cdf22b4d44cec7df97f3bfb00e9c1 | Rationally guided discovery platform for monoclonal antibodies against carbohydrate antigens using virus-like particle conjugate immunization and high throughput selection | NIH | MICHIGAN STATE UNIVERSITY | 5R21AI174001-02 | This project will establish a protein engineering platform for evolving monoclonal antibody binding affinity and specificity to solve the notorious challenge of developing clinical mAbs against tumor associated carbohydrate antigens (TACAs). Our central hypothesis is that the merger of Qβ carrier protein-elicited mAb discovery and rationally-guided directed evolution will outpace existing methodologies for discovering powerful antibodies against challenging TACA glycosylated biomarkers. TACAs are unique biomarkers to multiple tumor types, yet they have been underutilized for molecular imaging and diagnostics because of challenges in developing selective, potent binders. Distinct glycosylation patterns of tumor cell surfaces are hallmark features that arise during oncogenesis through changes in expression levels of glyco-processing enzymes. Problematically, these aberrant tumorigenic features are usually undetected by the immune system and rarely identified as non-self. Even when recognized as an antigen, weak binding against monovalent glycans leads to an insufficient immune response. To address this need, we will apply our directed evolution methodology to develop lead candidate mAbs against TACAs selective to cancer with in vivo binding of KD<10nM and specificity >100-fold binding above control cells. This will be accomplished by first generating a diverse panel of TACA-specific antibodies via immunization of transgenic mice with multivalent Qβ vaccines. Dominant antibodies will be isolated and characterized for paratope diversity and the ability to selectively bind the glyco-targets. Next, we use rationally- guided directed evolution to achieve mAb binding affinity and specificity. Multiple TACA-specific mAbs obtained through immunization will undergo high-throughput yeast display directed evolution with site-wise diversification based on structural, stabilizing, and phylogenetic factors to overcome the routinely low affinity of anti-carbohydrate binders. Specificity and affinity will be evaluated against multiple human tumor cell lines. This project will: 1) establish a platform that drastically reduces initial discovery time for translatable molecular imaging and diagnostic tools against carbohydrate antigens; 2) significantly advance understanding of tumorigenic cell glycosylation patterns; and 3) mark a major step towards improving sensitivity and specificity of biomarker-based diagnosis of cancers including ovarian, breast, and pancreatic cancers. Project narrative: Biomarker discovery and diagnostic options for aggressive diseases such as cancer are imperative for better disease outcomes. A platform for rapidly discovering and evolving novel antibodies against glycoprotein cancer biomarkers will be developed. This new strategy can help significantly enhance the treatment options available to cancer patients. | Non-SBIR/STTR | 6project_grants_public |
gen_cf2db1b0a36f138fbc6517e82c50320a | Mechanisms of action and therapeutic targeting of the CARM1-NFIB axis in small cell lung cancer | NIH | UNIVERSITY OF TX MD ANDERSON CAN CTR | 1R01CA272843-01A1 | ABSTRACT Treatment options for small cell lung cancer (SCLC) patients have remained largely unchanged for 3 decades, with no new FDA-approved treatments for 20 years, and no targeted therapies. We recently performed a screen for targets of an arginine methyltransferase called CARM1 and identified the NFI family of transcription factors as substrates for this PRMT. Importantly, NFIB harbors both oncogenic and metastatic promoting activities in the context of SCLC development. We confirmed that CARM1 functions as a transcriptional coactivator for NFIB. Based on these finding, we hypothesize that CARM1 methylation of NFIB is critical for its tumor-promoting functions. To further support this premise, we have generated a Nfib knockin mouse that harbors a R-to-K mutation in the CARM1 methylation site. When this mouse is crossed onto a SCLC genetically engineered mouse model (GEMM) the life expectancy of these mice is lengthened by a third (from 200 to 300 days), which is almost identical to the impact of Carm1-loss in the same GEMM. These finding raise the possibility of targeting SCLC with CARM1 small molecule inhibitors. We have also identified an effector molecule (TRIM29) for the CARM1 methylation site on NFIB. In this proposal we plan to: (1) perform a deep mechanistic analysis of this newly discovered CARM1/NFIB/TRIM29 signaling axis, and (2) investigate the therapeutic potential of targeting this axis using a battery of pre-clinical mouse models. PROJECT NARRATIVE We propose to investigate the mechanisms of action of the CARM1 methyltransferase in controlling small cell lung cancer pathogenesis via methylation of NFIB and therapeutic validation of small molecule inhibitors of CARM1 using pre-clinical cancer models. | Non-SBIR/STTR | 6project_grants_public |
gen_cb25cb342a613ecda4a0d0520da034c0 | Interferon regulation of gamma delta intraepithelial lymphocyte activation | NIH | ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI | 7R01DK119349-05 | PROJECT SUMMARY. Immune surveillance at mucosal barriers is essential to provide an immediate defense against invasive microbes, yet must also be tightly regulated limit the potential for autoimmunity. Intraepithelial lymphocytes expressing the γδ T cell receptor (γδ IEL) bridge innate and adaptive immunity, and function as a first line of defense by promoting mucosal barrier integrity. Recent reports demonstrate that basal γδ IEL function is influenced by extrinsic microbial signals. Although commensal-induced tonic type I interferon (IFN) signaling has been shown to prime mucosal innate immunity and host responsiveness to pathogen, the involvement of type I IFN in γδ IEL activation and epithelial surveillance remains unknown. Our preliminary data demonstrate that constitutive low level type I IFN signaling regulates the appropriate number and proportion of Vγ TCR subsets in the epithelial compartment and maintain these cells in an actively patrolling, yet immunologically quiescent state. We now show that impaired interferon α/β receptor (IFNAR) activation induces a dysregulated γδ IEL phenotype, characterized by hyperproliferation, hypermotility and enhanced IL-4 expression. Further, we find that pathogen-associated levels of type I IFN amplify γδ IEL effector functions, including epithelial surveillance. Therefore, we propose to interrogate the mechanism by which tonic type I IFN signaling maintains γδ IEL homeostasis, whereas amplification of type I IFN in response to pathogen enhances γδ IEL effector function. In the first aim, we will take advantage of genetic models that permit the inducible γδ T-cell-specific deletion of IFNAR to examine the role of tonic IFNAR/STAT signaling in the maintaining γδ IEL homeostasis through appropriate regulation of different Vγ subsets. We will also investigate the mechanisms by which IFNAR signaling regulates crosstalk between different γδ IEL subsets and how this influences the proliferation, motility and effector function of these cells. Next, we will determine the functional consequence of γδ IEL dysregulation on epithelial barrier integrity under steady-state conditions. In the second aim, we will examine the mechanisms by which type I IFN amplifies γδ IEL effector function following viral infection. Using the novel intravital microscopy techniques that we pioneered and our ability to move fluidly between in vitro and in vivo models, we will investigate the molecular signals induced by pathogen-associated levels of type I IFN to enhance γδ IEL epithelial surveillance and activation. Lastly, based on the protection conferred by γδ IELs in response to enteric pathogens, we will examine the role of type I IFN-induced γδ IEL activation in the context of acute enteric viral infection. By combining, temporal and cell-specific gene targeting, cutting edge live imaging techniques, and novel models to analyze γδ IEL function ex vivo, we expect to define the molecular mechanisms by which type I IFN regulates γδ IELs under homeostatic conditions and during infection. The proposed studies will provide new insight into the molecular mechanisms that regulate γδ IEL activation and the extent to which enhanced γδ IEL effector function affects epithelial integrity and host defense. PROJECT NARRATIVE. The proposed research is relevant to public health because it will elucidate the fundamental biological processes by which the immune system contributes to the mucosal barrier. This essential knowledge is required to understand mucosal surveillance during intestinal homeostasis and in response to pathogen infection. This work will therefore directly support the overall NIH mission of developing fundamental knowledge that will help reduce the burden of human disease and promote the NIDDK goal of improving digestive health. | Non-SBIR/STTR | 6project_grants_public |
gen_94f0bb2f3c0ec71f8268237bfaed0cbf | Dietary modulation of Paneth cells | NIH | WASHINGTON UNIVERSITY | 1R01DK136829-01 | ABSTRACT One of the underappreciated effect of obesity is the repression of gut innate immunity, as we recently showed in overweight/obese patients, using Paneth cell phenotype (as a surrogate for Paneth cell function) as a proof- of-concept. Wild type mice fed with western diet (WD) also developed Paneth cell defects. We further showed that WD consumption results in microbiota-mediated increase in deoxycholic acid (a secondary bile acid), which activates the FXR pathway in the ileum. FXR activation in the intestinal epithelium, as well as FXR- mediated type I interferon activation in myeloid cells, collectively trigger Paneth cell defects. The critical questions that need to be addressed before translating these findings to clinic include how individual dietary components that impact Paneth cell function, and if long-term WD consumption renders Paneth cell defects irreversible to dietary switch alone. Our long-term goal is to dissect the cellular and molecular mechanisms of how WD consumption affect the intestinal stem cell (ISC) and Paneth cell biology. These discoveries will facilitate design of trials for obese patients with gut innate immunity dysfunction. The objective of this proposal is to determine how WD consumption induces Paneth cell abnormality. The central hypothesis is that long-term consumption of dietary fructose result in Paneth cell defect due to diminished capacity of Paneth cell replenishment (by Paneth cells themselves and/or intestinal stem cells [ISC]). Our rationale is that identification of the mechanism(s) to restore Paneth cell function will offer new therapeutic opportunities for many patients, such as those with inflammatory bowel disease and graft versus host disease, of which Paneth cells play a critical role in pathogenesis. Our preliminary data suggest that dietary fructose consumption alone is sufficient to trigger Paneth cell defects, and that Paneth cell defects after long-term (≥12 months) WD consumption are not reversible by switching to standard diet. Our specific aims will test the following hypotheses: (Aim1) How dietary fructose directly induces Paneth cell defects; (Aim 2) Long-term WD consumption will impair the capacity of Paneth cells as well as ISCs to repair and replenish defective Paneth cells. Upon conclusion, we will understand the role for dietary fructose in modulating Paneth cell and ISC function. This contribution is significant since it will establish intestinal epithelial fructose catabolism and associated genes as therapeutic targets. The proposed research is innovative because we investigate how long-term exposure to poor diet on gut innate immunity, a heretofore-unexamined process. We also use state-of-the-art ISC culture system and scRNA-seq and scATAC-seq techniques to identify molecular and cellular targets that affect Paneth cell and ISC functions. Identifying the mechanisms of how diets regulate a key disease-relevant cellular phenotype will provide insight into other inflammatory disorders. PROJECT NARRATIVE: The proposed research is relevant to the public health because obesity, which is increasing in prevalence worldwide, represents a major national cost measured by both patient quality of life and economic burden. One under explored areas in obesity-related pathophysiology changes is the defects in mucosal innate immunity as we have previously shown, using Paneth cells as a proof-of-concept. Identifying the causes of how obesogenic diet reduces Paneth cell function will provide insight into the development of intervention strategies for diseases of which Paneth cells play a critical role, such as inflammatory bowel disease, graft versus host disease, and gut infection. Our preliminary data suggests that diet-induced obesity (due to western diet [WD] consumption) leads to Paneth cell dysfunction in both humans and mice by activating FXR signaling in the intestinal epithelium and myeloid cells. Upon conclusion, we will establish how dietary fructose (a key ingredient in WD) triggers Paneth cell defects, and determine the mechanisms by which Paneth cells themselves and intestinal stem cells repair defective Paneth cells in the context of long-term WD/HFrD consumption. Our data will help design intervention strategies for obese patients with Paneth cell defects. | Non-SBIR/STTR | 6project_grants_public |
gen_f4245b59d16be93506e9f43f768d7976 | Cancer-based discovery of novel mechanisms of chromatin control | NIH | ST. JUDE CHILDREN'S RESEARCH HOSPITAL | 5R01CA273455-02 | Project Summary/Abstract: The systematic sequencing of cancer genomes has revealed a high prevalence of mutations in genes encoding chromatin regulatory proteins. Of these aberrations, mutations in genes encoding subunits of SWI/SNF (BAF) chromatin-remodeling complexes are the most frequent, collectively occurring in over 20% of all cancers. Whereas most genes that are mutated at such high frequencies in cancer have been studied for many decades, recognition of a prominent role for SWI/SNF mutations is much more recent. The first link between SWI/SNF and cancer came when the gene encoding the SMARCB1 subunit was found to be biallelically inactivated in nearly all cases of the highly aggressive and lethal cancer of early childhood termed malignant rhabdoid tumor (RT). Notably, these RT cancers are genomically stable and diploid, rendering them a highly useful model in which to study the effects of SWI/SNF disruption. Our long-term goals are to elucidate the function of SWI/SNF complexes, to determine how their loss leads to oncogenesis, and to translate this understanding into novel highly effective therapies. Our group established SMARCB1 to be a bona fide and potent tumor suppressor and later made high-impact discoveries that help define mechanisms by which SWI/SNF mutations lead to dysregulated cell proliferation. Our findings to date suggest a model whereby SWI/SNF-facilitated control of transcription underlies cellular fate specification, with disruption of this control being the basis for cancer formation. We hypothesized that loss of SMARCB1, while driving cancer growth, also creates unique vulnerabilities. To identify such vulnerabilities, we undertook a rigorous screen involving 21 RT cell lines compared to 800 other cancer cell lines. From this screen, we identified and subsequently validated two novel genes as being required specifically and potently for RT cell survival. Using multiple approaches and tools, we have validated both genes as specifically essential in RT cells. Our subsequent preliminary data reveal that both genes have unanticipated novel roles in chromatin regulation: both regulate active chromatin and specifically facilitate acetylation of lysine residues on histone H3 that facilitate transcription of target genes. We now hypothesize that these genes cooperate with SWI/SNF complexes, and that elucidation of their function will provide novel insights into chromatin-mediated regulation of transcription, mechanisms by which mutation of SWI/SNF subunits drive cancer, and vulnerabilities created by SWI/SNF mutations. Additionally, both genes bring the opportunity for therapeutic targeting. In the proposed work, we will define the mechanistic relationship between these genes and SWI/SNF and determine the mechanism underlying the specific vulnerabilities in RT. Taken together, these efforts have potential for substantial impact by broadening understanding of the roles of chromatin regulators in normal cells and in transformation and by identifying highly specific new therapeutic targets for these lethal childhood cancers. Project Narrative: Mutation of genes involved in chromatin regulation frequently underlie cancer. We have identified and validated two genes that when deleted, cause cell death specifically in childhood cancers mutant for the SWI/SNF chromatin remodeler subunit SMARCB1. The proposed studies are designed to determine the normal function of these two genes and to define the mechanisms by which they kill these childhood cancers, with the ultimate goal of yielding new therapeutic approaches. | Non-SBIR/STTR | 6project_grants_public |
gen_0138e22ebc9004e75dc2f37c64b25384 | The effects of masculinizing gender-affirming hormone therapy for transgender men on susceptibility to HIV-1 infection modelled ex vivo in cervical mucosal tissue | NIH | UNIVERSITY OF ALABAMA AT BIRMINGHAM | 5R21AI178872-02 | PROJECT SUMMARY/ABSTRACT There are approximately 1.3 million transgender adults in the US, and about 467,000 of these individuals (~36%) are transgender men. Transgender men are individuals who were assigned female at birth but identify as male. Trans men may transition physiologically from female to male by receiving masculinizing hormone therapy and/or hysterectomy. Those in the trans male community participate in diverse sexual behaviors and lifestyles resulting in unique risks to STIs, especially HIV-1. Currently, there is a significant knowledge gap of the impact of HIV-1 on trans men, including limited knowledge regarding the effects of testosterone therapy on HIV-1 susceptibility and acquisition. Over 70% of trans men receive testosterone to promote masculine characteristics and reduce secondary female sex characteristics. Trans men treated with testosterone report symptoms of vaginal dryness and loss of elasticity, which increase mucosal tissue breaks, which contribute to increased risk of HIV-1 transmission in trans men. Trans men treated with testosterone for at least one year have significantly reduced levels of Lactobacillus comprising the vaginal microbiome, which correlates with bacterial vaginosis, and thus increased risk of HIV transmission. Like other androgens, testosterone is a steroid hormone that interacts with many different cell types, broadly affecting both innate and adaptive immunity through its effect on toll-like receptors, immune-response cells, and pro- and anti-inflammatory cytokines. Testosterone has broad-ranging effects on adaptive and innate immune functions and acts in a dynamic and often antagonistic manner with other androgens, particularly dehydroepiandrosterone (DHEA), to modulate the development and function of immune response cells. The central HYPOTHESIS of this research proposal is that testosterone alters cellular and immunologic responses in the cervical mucosa that affect susceptibility to HIV-1 infection. To interrogate this hypothesis, we propose to characterize certain cellular and innate immunologic properties of cervical mucosal tissue obtained from transgender men receiving gender-affirming masculinizing therapy, and undergoing medically indicated hysterectomies, and to correlate these findings to tissue susceptibility to HIV-1 infection ex vivo. We anticipate identifying specific alterations in the cervical mucosa that correlate with testosterone therapy and altered susceptibility to HIV-1 infection. If successful, our findings will provide new underpinnings for future hypothesis-driven research focused on HIV-1 prevention strategies for transgender men. The research proposed in this R21 grant application is guided by the following SPECIFIC AIMS: 1. Determine the effects of testosterone on the susceptibility of cervical explant tissue to HIV-1 infection and populations of T lymphocytes; and 2. Determine the effects of testosterone treatment on cytokine and chemokine expression in cervical tissue. PROJECT NARRATIVE This R21 grant application proposes to characterize the effects of gender-affirming testosterone therapy for transgender men on the cellular and innate immunological properties, and susceptibility to HIV-1 infection of cervical mucosa. Using an ex vivo model of HIV-1 infection in cervical explant tissue derived from medically indicated hysterectomies, we anticipate identifying specific alterations in the cervical mucosa that correlate with hormone therapy and increased susceptibility to HIV-1. If successful, our findings will provide vital underpinnings for future studies focused on HIV-1 prevention strategies for trans men. | Non-SBIR/STTR | 6project_grants_public |
gen_08fa50e713a6dde3970dc9f73004ff9f | Exploiting Carbon Monoxide Biofoams to Radio-Sensitize Rectal Cancer Cells While Protecting Normal Bowel | NIH | UNIVERSITY OF IOWA | 5K08CA276908-02 | ABSTRACT An estimated 45,230 individuals were diagnosed with rectal cancer in the United States in 2021. Rectal cancer is primarily managed using a combination of chemotherapy, radiation, and surgery. Surgical resection of the rectum is associated with long-term functional deficits and decreased quality-of-life. Therefore, strategies to improve response to neoadjuvant chemoradiotherapy could increase non-surgical curative rates and enhance quality-of-life for rectal cancer patients. Carbon monoxide (CO) at low, non-toxic concentrations has been shown to provide paradoxical anti-tumor effects while inhibiting inflammation and oxidative-stress induced normal tissue injury that could serve as an adjuvant treatment to enhance chemo-radiotherapy efficacy. CO is a product of heme catabolism regulated by the Nrf2 transcription factor and the cytoprotective gene Heme Oxygenase-1 (HO- 1). The biochemical mechanisms by which CO simultaneously sensitizes tumor cells to die while preserving normal cell survival are unknown but likely involve fundamental differences in oxidative metabolism between cancer and normal cells. Identifying targetable redox sensitive mechanisms underlying the activity of CO in rectal cancer could rapidly lead to translational therapeutic approaches for improving radiation responses in cancers while limiting normal tissue injury. We have developed exciting new methods for CO delivery through the gastrointestinal (GI) tract to overcome the challenges of inhaled CO. Using these GI formulations to deliver CO, our central hypothesis is that CO, delivered as a safe biofoam, selectively chemo-radio-sensitizes rectal cancer while reducing normal tissue injury. Further that the mechanism involves differential effects on Nrf2/HO-1 signaling and modulation of mitochondrial oxidative metabolism. We will evaluate the impact of cytoprotective CO biofoams on normal rectal tissue responses and oxidative damage after exposure to chemoradiotherapy and determine the effects of CO as an adjunct to therapy for rectal cancer in mice. NARRATIVE Rectal cancer patients have significantly improved quality of life when surgery can be avoided. Carbon monoxide, which has been shown to have beneficial properties at low concentrations with roles in modulation of diverse physiological and pathological processes such as inflammation and cancer, may improve anti-tumor response to chemoradiotherapy and paradoxically protect surrounding normal rectum. To improve neoadjuvant therapy and reduce the likelihood of requiring surgery, I propose the evaluation of novel CO biofoams as an adjunctive strategy to chemoradiotherapy and will evaluate the mechanism of protection in tumors and normal tissue. | Other Research-Related | 6project_grants_public |
gen_98e6076980628c308cf50088779c73cc | Molecular basis of plasma membrane rupture in lytic cell death and its inhibition by cytoprotective agent glycine | NIH | CASE WESTERN RESERVE UNIVERSITY | 5R35GM151043-02 | PROJECT SUMMARY/ABSTRACT Plasma membrane rupture (PMR) in lytic cell death—including pyroptosis, necroptosis, and post-apoptotic secondary necrosis—is a cataclysmic event that releases large-size intracellular molecules known as damage- associated molecular patterns (DAMPs), which in turn propagate the inflammatory response. Lytic cell death plays an important role in host defense against pathogen infections, but its dysregulation is also implicated in many inflammatory diseases and pathological conditions. PMR was thought to be a passive event, until a recent study identified NINJ1 to be responsible for carrying out this process. NINJ1 is a 16-kDa plasma membrane protein previously identified to be mediating cell adhesion through homotypic binding. It has two transmembrane helices and one extracellular amphipathic helix. NINJ1 undergoes oligomerization to induce PMR, and the amphipathic helix seems to play an important role in this process. However, a highly similar protein NINJ2 in the plasma membrane bearing a similar amphipathic helix does not induce PMR. To understand the molecular basis of NINJ1-oligomerization mediated PMR, we use cryogenic electron microscopy (cryoEM) to study the structures of NINJ1 and NINJ2 oligomers. The progress we have made is shedding light on a mechanistic understanding of this process. However, it also points to more hypotheses that need to be tested in order to fully understand this fundamentally important process. PMR is also linked to glycine cytoprotection in a very recent study. It was demonstrated that glycine treatment prevented NINJ1 oligomerization and thus prevented PMR in bone marrow derived macrophages receiving various forms of lytic cell death stimuli. We will include glycine treatment in our test of hypotheses for the search of signal that triggers NINJ1 oligomerization. This further elucidation of the molecular basis of glycine cytoprotection would inform the development of better cell preservation strategies or agents. PROJECT NARRATIVE Lytic cell death is a self-protection mechanism against pathogen infection or even cancer, but its dysregulation also results in various inflammatory diseases and tissue injuries. Plasma membrane rupture is the cataclysmic step in lytic cell death that further propagates the inflammatory responses. NINJ1 is a newly identified protein to be responsible for carrying out plasma membrane rupture, but its working mechanism is not clear. Glycine is a widely used additive in various ex vivo organ preservation solutions during organ transplantation for its cytoprotection effect. Glycine cytoprotection is recently demonstrated to be a result of inhibiting plasma membrane rupture. Our proposed studies aim to understand the structural basis of NINJ1-mediated plasma membrane rupture and its inhibition by glycine treatment. This research will have the potential of identifying novel therapeutic targets for various inflammatory diseases and informing the development of better cell preservation strategies or agents. | Non-SBIR/STTR | 6project_grants_public |
gen_a95c2c856af8aef1771d7e69e60c883a | Interactions between Trichuris and the gut microbiota | NIH | UNIVERSITY OF PENNSYLVANIA | 5R01AI179896-02 | PROJECT SUMMARY Parasitic nematodes colonize over a billion people worldwide and are associated with a range of maladies. Among these parasites, the whipworm Trichuris trichiura is particularly difficult to manage with existing medication that target adult worms due to drug resistance and frequency of reinfection. Following ingestion by the host, Trichuris eggs hatch in the gastrointestinal tract where they mature into adulthood and produce eggs, which are then deposited back into the environment in feces to perpetuate the cycle. As such, the entire time the parasite inhabits the host is spent in the presence of a diverse community of bacteria that are part of the gut microbiota. We and others have shown that colonization by Trichuris trichiura and the model parasite Trichuris muris restructures intestinal bacterial communities in humans and mice, respectively. Consistent with interactions between parasitic nematodes and the gut microbiota, bacteria are required for T. muris to complete its life cycle in mice due to a role for bacteria in promoting egg hatching. The details of how the gut microbiota mediate egg hatching and the reproductive fitness of Trichuris remain obscure. In preliminary results, we identified bacteria that differentially affect T. muris egg hatching and established a novel C. elegans screen to identify bacterial gene products that have a conserved role in the reproductive fitness of free-living and parasitic nematodes. The main objective of this proposal is to identify mechanisms by which bacteria mediate Trichuris egg hatching and affect subsequent colonization of the host. First, based on our findings that T. muris and C. elegans share a requirement for an arginine-dependent byproduct of E. coli, we will identify this metabolite and determine how it impacts host reproduction using the two nematode models. We will specifically test the hypothesis that polyamines are the key arginine-dependent factors in this system. Then, we will delve deeper into how bacteria-egg interactions mediate hatching by determining the role of chitinase produced by the parasite and compare bacterial species that promote versus impede hatching. Lastly, we will determine whether bacteria displaying superior egg-hatching activity enhance T. muris infection of mice, and using a new assay we developed for T. trichiura, extend our observations to the human parasite. We believe these innovative approaches will improve our understanding of how parasites have adapted to their host environment, and ultimately reveal vulnerabilities in the Trichuris life cycle that can serve as therapeutic targets. PROJECT NARRATIVE Parasitic intestinal worms and gut bacteria have co-evolved with humans and other mammals. A better understanding of how worms and bacteria interact in the gut may lead to new ways of controlling worm infections and reducing disease. | Non-SBIR/STTR | 6project_grants_public |
gen_fabbe23f9178a1c014c2ae5fcda78f48 | Exploration of the immunosuppressive function of RBMS3/PRRX1 axis in TNBC | NIH | WAYNE STATE UNIVERSITY | 1R21CA273771-01A1 | Abstract The epithelial to mesenchymal transition (EMT), a developmental process related to tissue repair and pathological processes, has been found to occur in the progression of carcinomas to invasive and metastatic disease. Accumulated evidence suggests the EMT could contribute to the immunosuppressive function of cancer cells. However, the underlying molecular mechanism linking EMT and immunosuppressive function in cancer remain largely unknown. To tackle this problem, our research group developed an integrative transcriptomic approach to combine expression profiling of breast cancer cell lines and several mammary epithelial cell EMT models. This screen identified RNA-binding motif single-stranded interacting protein 3 (RBMS3) as being significantly and reproducibly associated with EMT. We further showed that RBMS3 stabilized a group of EMT-related genes, including PRRX1. Functional analysis demonstrated the RBMS3/PRRX1 axis is responsible for maintaining mesenchymal status and motility properties of breast cancer cells, as well as controlling a group of pro- inflammatory cytokines. More importantly, knockdown of RBMS3 in TNBC MDA-MB231 cells results in a significant delay of tumorigenesis in vivo, which is not observed in vitro. These results indicate RBMS3 mediates breast cancer progression, potentially by simultaneously increasing invasive potential and promoting an immunosuppressed tumor microenvironment. In this study, we propose to investigate the effect of RBMS3/PRRX1 axis on immunosuppression and breast cancer progression in immunocompetent animal models and explore the potential impact of targeting RBMS3/PRRX1 axis in facilitating immunotherapy in TNBC models. We expect the proposed studies to be completed within two years, with two critical outcomes: 1) revealing the role of RBMS3/PRRX1 axis in driving TNBC progression through detailed analysis of the alterations of immune- microenvironment; 2) proof-of-concept evidence that targeting RBMS3/PRRX1 axis will facilitate immunotherapy for TNBC treatment. These results will lay a solid foundation for further development of specific targeting RBMS3/PRRX1axis for treatment of TNBC. By achieving these goals, we will be able to address the following overarching challenges: 1) identify why some breast cancers become life-threatening metastases; and 2) eliminate or reduce the mortality associated with metastatic breast cancer. Project Narrative In this proposal, we will employ complementary molecular, cellular, genetics, and state-of-the-arts next-generation sequencing techniques, as well as various mouse models, to investigate the function of RBMS3/PRRX1 axis in regulating immunosuppressive effect and response to immunotherapy for TNBC. The results of this study will not only provide insight into the mechanism of TNBC progression, but also lay the foundation for establishing the RBMS3/PRRX1 axis as a therapeutic target for TNBC treatment. | Non-SBIR/STTR | 6project_grants_public |
gen_c4874b6f3d7faf286594fad8ab58cea0 | Crosslinking-based targeted therapy for triple-negative breast cancer | NIH | ARIZONA STATE UNIVERSITY-TEMPE CAMPUS | 1R21CA280458-01 | Abstract The HER1 and/or HER3 receptors are overexpressed in most triple-negative breast cancers (TNBCs), a subtype of breast cancer that lacks estrogen receptor, progesterone, and HER2 expression and is associated with poor prognosis and a high lung and brain metastasis rate. Chemotherapy is the mainstay of TNBC treatment. Our long-term goal is to develop more effective therapy for this aggressive subtype of breast cancer. In this proposal, we aim to develop a novel crosslinking-based targeted therapy. For this purpose, we will create a series of multi-functional DNA-affibody-drug nanoparticles containing multiple copies of HER1 and/or HER3 specific affibody molecules covalently coupled to a DNA nanostructure, of which the latter binds to tens of small molecule drugs such as THZ1. In the presence of these nanoparticles, HER1+ and/or HER3+ TNBC cells will be tightly bound together, thus preventing metastasis in its initial stage. In the cross-linked TNBC cell clusters, the small molecule anticancer agent THZ1 reversibly bound to the nanoparticles will be released slowly, resulting in killing of crosslinked TNBC cells. These nanoparticles can bind to HER1+ and/or HER3+ TNBC cells in primary tumors, metastatic sites, and circulation. In our preliminary study, cultured HER1+ TNBC cells were crosslinked together to form cell clusters with a 96% crosslinking efficiency, 99% migration inhibition, and 90% invasion inhibition by a DNA-4ZHER1-THZ1 nanoparticle (drug-to-cargo ratio is 50). To enhance the efficiency and overcome the resistance to HER1 inhibitors, a novel DNA-2ZHER1-2ZHER3-THZ1 nanoparticle that targets dual HER1 and HER3 will be synthesized and tested in this study. Our design and hypothesis will be tested with the following specific aims: In specific aim 1, we will prepare a DNA-2ZHER1-2ZHER3- drug nanoparticle for targeting HER1- and/or HER3-overexpressing TNBC. In specific aim 2, we will evaluate the inhibition of HER1- and HER3-overexpressing TNBC cell growth and metastasis by the DNA-2ZHER1-2ZHER3- drug nanoparticle in vitro. In specific aim 3, we will study distribution and pharmacokinetics of selected DNA- 2ZHER1-2ZHER3-drug nanoparticle in healthy mice, and to evaluate its suppression of mammary tumor growth and metastasis in TNBC xenograft models. If successful, this innovative approach will open a new avenue for developing new therapy for treatment of TNBC. The pilot study will lay a foundation for further studies to investigate the utilities and mechanisms of the new class of anticancer agents for targeted therapy. Project Narrative In this proposal, our long-term goal is to develop a novel “crosslinking-based targeted therapy” for treatment of triple-negative breast cancer patients. If successful, this innovative therapy for the prevention and treatment of TNBC metastasis should have a significant impact on reducing the mortality rate in triple-negative breast cancer patients. | Non-SBIR/STTR | 6project_grants_public |
gen_1ef4c00da2f313df83922900cacdecd5 | Neuronal Regulation of Skeletal Development and Repair | NIH | UNIVERSITY OF MARYLAND BALTIMORE | 7R01DE031028-08 | This is a renewal application of a program investigating the role of sensory nerves in bone. Our studies during the first funding period demonstrate that NGF-dependent TrkA signaling by sensory nerves is the primary driver of angiogenesis and osteogenesis in the developing femur and skull. In these avascular settings, acute up-regulation of NGF in mesenchymal lineage cell domains is followed by nociceptive fiber ingrowth, which subsequently home to locations of proliferating mesenchymal cells. Blockade of sensory nerve ingrowth, either by inhibition of TrkA signaling or disruption of NGF, retards vascularization and disrupts femoral and calvarial bone formation. Histological data in the calvaria model revealed that loss of sensory nerve fibers is associated with reduced numbers of proliferating osteogenic precursors in the sutures and premature suture closure. These observations suggest a paradigm in which sensory nerves function in developing bone to maintain mesenchymal stem cell plasticity, a concept well established in models of limb regeneration and supported by recent studies in developing mouse femur. Our preliminary findings directly examining the interaction of sensory nerve axons with MSCs in microfluidic chambers suggest that infiltrating DRG nerve fibers induce MSC proliferation, but limit differentiation in a non-contact dependent fashion. These effects are accompanied by upregulation of osteoprogenitor mitogens (e.g. TGF) and inhibitors of MSC differentiation (e.g. follistatin-like 1). Together, this data support the premise that TrkA+ sensory nerves function in developing bone to maintain stem cells in a proliferative, undifferentiated state by delivering soluble factors that activate mitogenic and anti-differentiation signaling pathways. This conceptual model will be explored in studies divided into three Specific Aims. Specific Aim 1 will define the spatiotemporal patterning of TrkA+ skeletal sensory nerves in the developing cranium, and determine their influence on osteoprogenitor proliferation and cellular fate. Specific Aim 2 will identify key target genes in MSCs impacted by sensory nerve signals using previously validated co-culture methods. Specific Aim 3 will identify sensory axon-derived factors that regulate MSC proliferation and cell fate decisions. Our results should provide new insights into the fundamental roles sensory nerves play in skeletal morphogenesis, homeostasis and repair, and provide critical insight into the neuropathological manifestations associated with bone disorders in humans. PROJECT NARRATIVE Anyone who has broken a bone knows of the existence and function of skeletal sensory nerves. However, little is known regarding the function of these nociceptive fibers beyond the transmission of pain signals. In this project, we continue to characterize how skeletal sensory nerves spatially approximate osteogenic precursors in the developing bone to regulate stem cell expansion and faithfully pattern the mammalian skeleton. | Non-SBIR/STTR | 6project_grants_public |
gen_84e77cb31d319846d8947c856abbe42e | More Outside Your Door (MOYD) | NIH | ALASKA NATIVE TRIBAL HEALTH CONSORTIUM | 5R01HL168853-02 | ABSTRACT The Rural Alaska Community Action Program (RurAL CAP), which administers Head Start schools in rural Alaska, reports that 68% of Alaska Native children ages 3 to 5 years are overweight and 43% are obese. Diet, physical activity, and sleep hygiene are primary risk factors for childhood obesity, a risk factor for cardiometabolic diseases later in life. Notably, subclinical cardiovascular pathology can be detected as early as 3 years of age. Once living off the land and sea, Alaska Native (AN) people have suffered decades of colonization and in-migration, which have interfered with traditional food consumption, signaling a striking change in local food systems. Diet and physical activity are prime targets for early intervention, as taste preferences and exercise habits established in childhood forge lifelong adherence. Thus, we have designed More Outside Your Door (MOYD), a multilevel intervention, which at the individual level builds on our prior efforts to promote traditional diet and activities to decrease risk for obesity in AN preschoolers. At the family level, MOYD provides feedback to parents about their child’s health screenings; at the school level, it features traditional foods, outdoor traditional physical activities, and a culturally centered curriculum. At the community level, it offers resources to improve the built environment for safe outdoor play that invites all community members to participate. In partnership, the Alaska Native Tribal Health Consortium, RurAL CAP, and collaborators will refine and test the effectiveness of MOYD at 12 Head Start preschools in the Yukon-Kuskokwim region: (1) we will use qualitative methods with Head Start staff, parents, and community Elders to optimally tailor MOYD for southwest Alaska communities; and (2) we will conduct a stepped-wedge group-randomized trial to test its effect on children’s diet, physical activity (PA), body mass index (BMI), and other risk factors for obesity-related chronic disease over three years. Primary outcomes are diet (classroom meals, actigraphy-based physical activity, and body mass index. Secondary outcomes include individual child level measures of hemoglobin levels and educational and developmental progress; parent report of diet, activity, and sleep at home; and measures of traditional foods/sugar-sweetened/processed foods by carbon and nitrogen stable isotope ratios using fingernail clippings. We will also measure outcomes at the school level (classroom meals, outdoor time) and community level (objective data on use of play space). (3) We will conduct a process evaluation using the RE-AIM framework. Our Specific Aims are to (1) conduct preschool staff and parent/elder workgroups to refine the MOYD intervention for optimal effectiveness for obesity prevention among AN preschool children; (2) test the effectiveness of MOYD on a) diet, PA, outdoor time, and BMI; and b) improving other clinical and behavioral risk factors for obesity-related chronic disease in preschool children; and (3) implement the RE-AIM framework to evaluate dissemination and implementation of the MOYD intervention in Head Start programs. PROJECT NARRATIVE The preschool age is a crucial period of growth and an optimal time to begin to establish healthy eating and physical activity habits leading to better food and activity choices into adulthood, thereby minimizing risk for obesity-related diseases and decreasing the cardiometabolic disparities in this Indigenous population. More Outside Your Door is a multi-level, randomized, stepped-wedge intervention trial designed to reduce the disparity of childhood obesity in Yup’ik Alaska Native children by increasing the proportion of nutrient-dense traditional and traditional-like foods consumed and increasing physical activity, particularly outdoor activities related to traditional Yup’ik subsistence and lifestyle practices. This 5-year intervention trial targeting 3-5 year olds is conducted in partnership with Rural Action Community Action Program Head Start programs in 12 rural Alaskan communities, where each site is assigned annually to a wedge group to receive either a community-altered culturally-tailored traditional foods and activities curriculum intervention or the standard regional Head Start program intervention. | Non-SBIR/STTR | 6project_grants_public |
gen_43149b28df6c3455d33bed929067e73b | Improving awareness of women with hypertension: ROAR (Rural, Obese, At Risk) Career Enhancement Core | NIH | AUGUSTA UNIVERSITY | 5U54HL169191-02 | More than 1 in 3 women are living with heart disease and many are unaware of the risks. Hypertension is a major modifiable risk factor for cardiovascular disease (CVD), and ~47% of adults in the US have hypertension. In ~85% of cases, the cause of hypertension is unknown and only ~50% of patients taking medication achieve blood pressure (BP) control to recommended levels. A critical barrier to limiting premature death from CVD is lack of awareness surrounding the risks of CVD. The Career Enhancement Core (CEC) is designed to develop a comprehensive educational platform on SABV in CVD to increase awareness and education of CVD and the risks of uncontrolled hypertension in women across the health span through community outreach. The CEC is critical to the success of ROAR. The goals of the CEC are to develop a foundation for long-term relationships among the research, medical and lay communities in GA, MS and SC, and bring greater awareness to the risk of CVD in women and the consideration of SABV in the control of hypertension through education, mentorship, strategic partnerships, and training of community-engaged researchers. The ROAR-CEC has 3 objectives. Objective 1 is to develop a culturally diverse research pipeline of biomedical scientists trained in the field of SABV in CVD. This Objective will provide research career enhancement opportunities in SABV research, train a diverse scientific workforce, and promote mechanistic studies in SABV at different career stages. Objective 2 is to establish a community outreach pipeline of scientists trained to increase awareness of SABV and promote optimal cardiovascular healthcare for men and women. ROAR-CEC will leverage and strengthen existing community partnerships to engage in outreach to promote awareness about cardiovascular risk to men and women at local schools, colleges and HBCUs. Objective 3 is to empower the community to self-monitor and identify barriers to improving access to healthcare. This will be accomplished through innovative educational materials and qualitative assessment of barriers. ROAR-CEC takes a comprehensive approach to training and education in SABV in CVD and increasing community awareness of the risks of CVD. Our interdisciplinary team is uniquely prepared to address these important issues in at-risk populations in the Southern US. The impact of ROAR-CEC will be to train the next generation of scientists engaged in research related to women’s health and SABV, to change common perceptions and biases regarding women’s health and hypertension risk factors, and support the development of a new, self-aware, sustainable, and motivated community of researchers, educators, physicians and community members. ROAR will create synergy that will transform academic, clinical and community awareness of the importance of cardiovascular health in women. Approximately 19 million deaths were attributed to cardiovascular disease (CVD) globally in 2020, an increase of ~20% from 2010, and heart disease remains the leading cause of death for men and women in the US. The Career Enhancement Core (CEC) is designed to develop a foundation for long-term relationships among the research, medical and lay communities in GA, MS and SC to bring greater awareness to the risk of CVD in women and the consideration of SABV in CVD research. This will be accomplished through education, mentorship, strategic partnerships, and training of community-engaged researchers to increase research and awareness in at-risk communities throughout the Southeast. | Research Centers | 6project_grants_public |
gen_4591cf848fecfb4555b5ce90ed8ca212 | Mechanisms of Telomere Cohesion | NIH | NEW YORK UNIVERSITY SCHOOL OF MEDICINE | 1R35GM149355-01 | Project Summary/Abstract Telomeres, the specialized structures at chromosome ends, are comprised of TTAGGG repeats, telomere repeat containing RNA (TERRA), and the shelterin protein complex. Sister chromatids are held together from the time of their replication in S phase until their separation in mitosis by cohesin rings. Ring-mediated cohesion is essential to ensure accurate distribution of chromosomes to daughter cells in mitosis. Cohesion between sisters is also important for recombination and repair, particularly at repetitive sequences like telomeres, where it keeps them aligned. For this, more intimate contacts (in addition to the cohesin ring) are needed and as such, there are telomere-specific requirements for cohesion. Establishment of cohesion at telomeres requires shelterin subunits and associated proteins. Resolution of cohesion between telomeres requires the PARP, tankyrase. Tankyrase localizes to telomeres by binding to the TTAGGG-repeat binding shelterin subunit TRF1, in late S/G2 to resolve cohesion. In tankyrase-depleted cells sister telomeres remain cohered in mitosis despite normal resolution of arms and centromeres. This persistent telomere cohesion is not just an aberrant state induced by depletion of tankyrase, it occurs naturally in certain human cell types that lack telomerase and have critically short telomeres: normal aged cells and cancer ALT cells. Unexpectedly, (and shown by our lab in the last few years) this persistent cohesion is beneficial to cells; it serves a protective role to prevent premature senescence in aged cells and growth arrest in ALT cancer cells. The goal of our research for the next five years is to elucidate the proteins and mechanisms required for establishment and resolution of cohesion. We will build on our previous work where we identified shelterin subunits and associated factors that are required for cohesion and we will focus on our most recent discovery indicating a role for RNA (TERRA and TERRA R-Loops) in telomere cohesion. We will determine how the telomeric components contribute to the establishment, maintenance, and resolution of telomere cohesion in normal human cells, aging, and cancer. For cohesion establishment, we will determine how and when the required proteins are deposited on telomeres and how far into the chromosome they extend. For cohesion resolution, we will determine how recruitment of tankyrase serves to resolve cohesion. We will characterize the proteins that tankyrase itself recruits to telomeres and investigate how tankyrase uses functional compartmentalization to orchestrate the resolution process. We will determine how TERRA RNA and TERRA R-loops contribute to telomere cohesion in normal cells and to persistent telomere cohesion in pathological conditions of aging and ALT cancer. And finally, we will elucidate the full proteome of the cohered telomeric state. Ultimately, a complete understanding of telomere cohesion will elucidate fundamental mechanisms of chromosome stability and genome integrity in normal human cells and provide insights into prevention of premature senescence in aging cells and into promotion of premature death in ALT cancer cells. Project Narrative Telomere shortening drives genome instability in aging and cancer. The shortening process is tightly monitored to prevent a DNA damage response and premature cell cycle arrest. Mechanistic understanding of this process and the role of RNA will provide insights to prevent premature senescence in aging cells and to promote premature death in ALT cancer cells. | Non-SBIR/STTR | 6project_grants_public |
gen_1466bd22dd47a95f0ef63468efe86890 | Sex determination and the sex-determining locus in aedine mosquitoes | NIH | VIRGINIA POLYTECHNIC INST AND ST UNIV | 5R01AI179056-02 | Abstract Sex is critical to the survival and evolution of sexually reproducing organisms including mosquitoes. A dominant male-determining factor (M factor) is the primary signal that controls sex-determination in mosquitoes. Nix, the M factor in the yellow fever mosquito Aedes aegypti, is the first M factor found in mosquitoes. In a simplified model, the expression of Nix, a predicted RNA-binding protein, leads to male- specific splicing of the pre-mRNAs of two conserved transcription factors, doublesex (dsx) and fruitless (fru), which program male sexual differentiation. In Aedes the M factor is located within the male- determining locus (M locus) on one of the “autosomes”. This pair of “autosomes” are so-called homomorphic sex chromosomes that are cytologically indistinguishable except in the region around the sex locus. The Ae. aegypti M locus is a ~1.3 Mbp repeat-rich region that contains Nix and four other protein-coding and 25 long non-coding RNA genes. The Nix transgene alone, in the absence of the M locus, is sufficient to convert females into fertile albeit flightless males, and myo-sex, a myosin heavy chain gene also in the M-locus, is required for male flight. The M- and m-bearing chromosomes in Aedes mosquitoes provide an opportunity to gain insights into the evolution of homomorphic sex chromosomes. In addition to its basic biological importance, Ae. aegypti is a major vector for the dengue, chikungunya, and Zika viruses. No specific treatment for dengue exists and the first dengue vaccine is recommended only for a limited population. Prevention of these vector-borne infectious diseases relies heavily on effective vector control. However, increasing insecticide-resistance poses a significant threat. Therefore, novel control strategies are urgently needed. Only female mosquitoes feed on blood and transmit pathogens, and for the most part females determine the size and distribution of the mosquito population. We are interested in deciphering the mechanism of sex-determination, investigating sex chromosome evolution, and translating such fundamental knowledge into safe, efficient, and diverse methods to control diseases that are transmitted by Ae. aegypti. Building on recent progress, we will pursue the following specific aims: 1) Decipher the sex locus in Ae. aegypti, 2) Identify and characterize the target(s) of Nix, and 3) Develop efficient sex-separation methods through Nix-mediated innovations. Project Narrative Aedes aegpyti is a major vector of dengue, Zika, and chikungunya. We explore the sex- determination pathway and the sex-determining locus to develop new ways to separate the non- biting males from females to help control mosquito-borne infectious diseases. | Non-SBIR/STTR | 6project_grants_public |
gen_81a536db755b37f71bb9c5b841f34fde | Identifying the Structural Adaptations that Drive the Mechanically Induced Growth of Skeletal Muscle | NIH | UNIVERSITY OF WISCONSIN-MADISON | 5R01AR082816-02 | Project Summary / Abstract Mechanical signals play a major role in the regulation of skeletal muscle mass, and the maintenance of muscle mass contributes significantly to disease prevention and quality of life. Although the link between mechanical signals and the regulation of muscle mass has been recognized for decades, the mechanisms that control this process remain ill-defined. For instance, most studies indicate that the mechanically induced growth of skeletal muscle is driven by an increase in the size of the existing myofibers rather than an increase in the number of myofibers. Moreover, current models assert that the increase in myofiber size is mediated by an increase in the balance between the rates of protein synthesis and protein degradation which, in turn, leads to the accumulation of newly synthesized proteins (NSPs) and the concomitant structural changes that drive the growth response. For instance, it is well known that an increase in mechanical loading can lead to microstructural changes such as the radial growth of myofibers. Surprisingly, however, the ultrastructural adaptations that drive these microstructural changes have not been defined. Indeed, a number of foundationally important questions such as whether the radial growth of myofibers is driven by an increase in the size and/or the number of myofibrils have not been answered. Likewise, the location(s) in which NSPs accumulate during mechanically induced growth (i.e., the sites of growth) are not known. As such, one of the major goals of this project is to fill these gaps in knowledge. Another major goal is to develop a better understanding of the signaling events that control the different aspects of mechanically induced growth. For instance, our previous work has established that signaling through mTORC1 plays a central role in the process via which mechanical stimuli induce the radial growth of myofibers. However, our preliminary data indicate that the longitudinal growth of myofibers can also make a substantive contribution to the mechanically induced accretion of muscle mass, yet, unlike radial growth, the longitudinal growth of myofibers does not appear to require signaling by mTORC1. In other words, our preliminary data suggest that the radial and longitudinal growth of myofibers are regulated by distinct signaling pathways. Specifically, we propose that the radial growth of myofibers is driven by a mTORC1-dependent mechanism that we have coined as the “myofibril expansion cycle”, whereas the longitudinal growth of myofibers is mediated by a mTORC1-independent mechanism that involves transverse Z-line splitting of sarcomeres at regions called sphenodes. To test the validity of these hypotheses we will use advanced imaging techniques, various genetic interventions, two complementary models of mechanical load-induced growth, and our new state-of-the-art technology that enables us to visualize and quantify (with ≤10 nm resolution) where NSPs accumulate. Collectively, it is anticipated that the outcomes of this project will not only fill major gaps in our understanding of how mechanical stimuli regulate muscle mass, but they will also build the framework for future studies that are aimed at developing a better understanding of this highly important process. Project Narrative This project is relevant to public health because the outcomes will facilitate studies that are aimed at developing therapies that can attenuate or reverse the loss of skeletal muscle that occurs during a variety of conditions such as prolonged bed rest, cachexia, muscular dystrophies, myopathies, immobilization, and aging. | Non-SBIR/STTR | 6project_grants_public |
gen_c1578cab1a7fe8244a076a1b1bee63ba | Functional consequences of evolutionary innovation in histone repertoires | NIH | FRED HUTCHINSON CANCER CENTER | 5K99GM149928-02 | PROJECT SUMMARY Histone proteins package DNA into chromatin and regulate all DNA-templated biological processes in eukaryotes. Consistent with their essential function, mutations or misregulation of histones result in many diseases. While core histones primarily function in genome packaging, histone variants can replace canonical histones at unique genomic locations for specialized roles such as DNA damage response (DDR) or gene expression. Despite their essential functions, histone repertoires have undergone distinct lineage-specific changes. Such evolutionary novelty via gene fusions, duplications or sequence divergence is unexpected in conserved protein families and suggestive of an adaptive advantage for sequence innovation. This proposal will use evolutionary innovations in eukaryotic histone H2A repertoires to investigate the causes and consequences of evolutionary turnover of histone proteins. The most common eukaryotic H2A repertoire is made up of core histone H2A, and histone variants H2A.X and H2A.Z that are involved in DDR and gene regulation, respectively. However, two evolutionary shifts, both likely selectively advantageous, have occurred in yeast and Drosophila H2A repertoires. In yeast, H2A.X, entirely replaced canonical H2A, likely improving DDR and affecting processes like meiosis that depend on DDR. In Drosophila, H2A.X fused with H2A.Z giving rise to a unique H2Av variant. This fusion enriches DDR at heterochromatin which potentially restricts DDR- based transposition events to gene poor regions. To identify functional consequences of these evolutionary innovations, the applicant will re-engineer the ancestral eukaryotic H2A repertoire in yeast and flies. Specifically in S. cerevisiae, the applicant will engineer a core H2A and two variants H2A.X and H2A.Z, preventing H2A.X from being the core histone (Aim 1). In D. melanogaster, the fusion histone H2Av will be separated into H2A.X and H2A.Z uncoupling DDR and gene regulation functions (Aim 2). Changes to organismal chromatin packaging, and relevant biological functions including DNA repair, meiosis, fertility, and transposition will be interrogated. In Aim 3, the applicant will apply tools learnt in Aims 1 and 2 to study divergence of eukaryotic core H2A. Histones have tail sequences which are heavily post-translationally modified and play crucial roles for higher order chromatin packaging and protein interactions. The high sequence divergence across eukaryotic histone tails suggests that tails could facilitate unique lineage-specific functions. By engineering different tail sequences in yeast and flies, Aim 3 will reveal changes to chromatin packaging, and changes to processes such as mating, and quiescence in yeast, and fertility and development in flies. To launch this work, the applicant requires training in genetics, genomics, and phenotypic assays in two model organisms, yeast and flies. By leveraging, her expertise in evolutionary analyses and the power of well-established tools in yeast and flies simultaneously, in the future the applicant will study the biological basis and consequences of histone innovation including her own previous discoveries and co-evolving mechanisms. PROJECT NARRATIVE Eukaryotic histone proteins are fundamental to all biological processes and were historically thought to be highly conserved. However, histones proteins can be subject to evolutionary turnover in the form of gene duplications and dramatic sequence divergence in multiple eukaryotic lineages suggesting adaptive advantages. This proposal uses yeast and flies to investigate the fitness advantages of unique eukaryotic histone repertoires for two crucial functions – genome regulation and DNA damage response. | Other Research-Related | 6project_grants_public |
gen_fd2d14632315590fcfb68ea35ab7e621 | Multi-site Gastrointestinal Cancer Detection by Stool DNA Methylation | NIH | MAYO CLINIC ROCHESTER | 4R37CA214679-06 | PROJECT SUMMARY/ABSTRACT: Gastrointestinal (GI) malignancies lead cancer deaths worldwide, killing ~3 million annually. In the U.S., only colorectal cancers (CRC) are screened. Other GI cancers are not screened due to lack of accurate tests or because prevalence is deemed too low for cost-effective screening. Consequently, most patients with GI cancers present at late-stage and cure rates remain abysmally low. Effective early detection is desperately needed to improve outcomes. Our group was central in development and validation of the FDA-approved multi- target stool DNA test for CRC screening. We have begun to expand this approach, showing feasibility to detect supra-colonic GI cancers by stool DNA testing. However, it is critical for a non-invasive molecular test to localize the site of a primary cancer (“site-prediction”). Key preliminary data suggest that this may now be possible. First, we have completed rigorous next-generation sequencing to identify differentially methylated regions (DMRs) which appear highly discriminant for universal and site-specific detection of GI cancers. Second, we prioritized these DMRs by strict filtering criteria and performed a confirmatory study with statistical cross-validation on an independent set of CRC, gastroesophageal and pancreatico-biliary cancer, and normal control tissues. This showed that a panel of 8 selected DMRs could distinguish cancer from normal (95% accuracy) and assign organ site (94-95% accuracy for each category) with overall site-prediction accuracy of 88%. Third, these findings have been confirmed with novel DMRs assayed from stool specimens obtained from CRC and pancreatic cancer patients and normal controls (30, each). Using a 2-stage analysis, cancers were distinguished from controls at 90% specificity in the first stage. At stage 2, the markers accurately classified CRCs from pancreatic cancers with 90% accuracy. It is now our central hypothesis that luminal and ductal adenocarcinomas can be detected and localized by stool assay of universal and site-specific DMRs. This raises 3 key questions: 1) will stool assay of our novel DMRs show the high overall cancer sensitivity and the site-prediction we have seen in preliminary data; 2) will the DMRs be specific for cancer across a wide patient demographic spectrum and in the setting of non-malignant GI diseases; and 3) can sensitivity and specificity be improved by novel assay technology? These will be addressed in the following parallel, integrated, but independent specific aims: 1) Assess panel sensitivity and site-prediction accuracy in stool specimens for adenocarcinoma at esophageal, pancreatic and colorectal sites; 2) Confirm and evaluate DMR specificity in stool; and 3) Optimize novel assay conditions and marker selection for cancer detection and site-prediction at esophageal, pancreatic and colorectal sites. With our team’s strong track record, extensive stool archive, and unique access to a state-of-the art assay platform, we expect to demonstrate in a cost-efficient manner the feasibility of a novel DMR panel for the detection and site prediction of specific GI adenocarcinomas. Results will inform designs of future studies ranging from large- scale case-control studies (phase 2) on early-stage cancer and pre-cursors to pivotal cohort validation (phase 4) of a non-invasive multi-GI cancer screening test. The potential impacts on cancer control are far-reaching. PROJECT NARRATIVE: Public Health Relevance: The aggregate incidence of GI cancers across the whole digestive tract is the highest of any organ system; re-imagining the GI tract as a single target organ provides the rationale to screen the most lethal GI cancers simultaneously. Biomarkers that discriminate GI cancers by anatomic site make this approach practical by efficiently directing the diagnostic evaluation of positive test results. The project is relevant to NIH mission by applying knowledge that might extend healthy life and reduce the burden of illness due to GI cancers. | Non-SBIR/STTR | 6project_grants_public |
gen_ee4f459cfdc93d5b4ffb4ed939f04d91 | Determining the Functional Significance of Mutations Observed in Envelope Protein Following Serial In Vivo Passaging of Human-Simian Immunodeficiency Virus | NIH | TEXAS BIOMEDICAL RESEARCH INSTITUTE | 1R03AI174973-01A1 | Abstract/Summary: Commonly used animal models of HIV-1 include infection of macaques with Simian Immunodeficiency Virus (SIV) or Simian-Human Immunodeficiency Virus (SHIV) containing HIV envelope (Env) or reverse transcriptase. These animal models have been extremely useful in understanding HIV pathogenesis and disease progression, as well as understanding the efficacy of vaccines and drugs. However, the genetic difference between HIV-1 and SIV, and the absence of other HIV-1 genes such as gag, pol, vif, vpr, and nef in SHIV limits the utility of these models in vaccine studies. Ideally, good animal model of HIV-1 infection/AIDS would be infection of macaques with HIV-1. However, HIV-1 does not replicate in macaque cells due to the presence of retroviral restriction factors. HIV-1 can be made to replicate by substituting its accessory genes with SIV genes such as vif, vpx, vpr, and nef, which can counteract interferon-induced restriction factors in macaque cells. Human-Simian Immunodeficiency Virus (HSIV) is an HIV-1NL4-3 derivative with SIV vif gene substitution (named HSIV-vifNL4-3) that can replicate persistently in pigtail macaques (PTMs). However, infection did not result in high peak viremia and setpoint viral loads as observed during SIV infection of macaques. Serial in vivo passaging in PTMs was performed to enhance infectivity or replicative capacity of HSIV. Three rounds of animal-to-animal transfer of infected blood in 3 immunocompetent PTMs with starting initial inoculum containing a mixture of CXCR4- (HSIV- vifNL4-3 recovered from previously infected macaque) and CCR5-tropic HSIV (HSIV-vif derivative based on pNL- AD8 and Bru-Yu2) was conducted to generate pathogenic variants. Interestingly, all the macaques showed peak viremia close to or above 105 copies/ml and virus replication persisted for more than 20 weeks. Following in vivo passaging, three infectious molecular clones (IMCs) were recovered from passage 3 macaque (HSIV-P3 IMCs). Sequencing of HSIV-P3 IMCs showed several interesting mutations throughout the genome, perhaps suggesting adaptation to PTMs. These mutations could help the virus in overcoming restriction factors, or better utilization of host dependency factors, or they could help the virus escape host immune responses. Focus of this grant application is to determine the functional significance of mutations observed in envelope gene. The results from this study will provide valuable insights into the role of envelope gene in cross-species transmission of HIV-1 to pigtailed macaques. Project Narrative: We conducted in vivo passaging of Human-Simian Immunodeficiency Virus (HSIV) in pigtailed macaques and recovered infectious molecular clones (IMCs) from the passage 3 macaque. We have observed several non- synonymous mutations throughout the genome of recovered IMCs, perhaps suggesting adaptation to pigtail macaques. In this grant application, we propose to determine the functional significance of mutations observed in envelope gene of recovered IMCs. | Non-SBIR/STTR | 6project_grants_public |
gen_d58a697e74101b11fa1e3fbafee6557e | Impact of Obesity on Chemotherapy-Induced Cytotoxicity: Immune Cells and Skeletal Muscle | NIH | UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA | 1K99CA276891-01 | PROJECT SUMMARY / ABSTRACT The number of cancer patients and cancer survivors continues to increase while the prevalence of obesity also continues to increase in the US. Obesity is associated with a greater risk for developing 40% of cancers and two of the four most prevalent cancers (i.e. breast and colon), are tightly linked with obesity. 5 fluorouracil (5FU) remains the first line of treatment for colon cancer despite 5FU’s well established toxicities - cytopenia, mucositis, anorexia, weakness, and fatigue. These toxicities contribute to reduction in relative dose intensity, increase patient susceptibility to infection, and lead to debilitating functional impairments that not only burden the patient, but also the patient’s support system. Given the increased prevalence of obesity in the US, it is increasingly more likely that those needing to undergo anti-cancer treatment will be obese. While it is common practice to apply a dosing cap, the current recommendations for the treatment of obese cancer patients are to give full body surface area dosing regimens, despite some evidence suggesting obese patients have exacerbated drug toxicities and reduced survival. This evidence is not ubiquitous as certain investigations have highlighted better prognosis and survival with increasing BMIs. I have discovered that obese mice are unable to sustain 2-3 cycles of 5FU. This I have attributed to a reduction in dihydropyrimidine dehydrogenase (DPD), the enzyme responsible for catabolizing 5FU in the liver. This has highlighted the need for mechanistic inquiry into the impact of obesity on 5FU’s toxicities; my K99/R00 proposal addresses this unmet need and will serve as a platform to launch my independent career in this domain. The overall goal of my proposed K99/R00 is to: 1) to understand the impact of obesity on 5FU’s anti-cancer efficacy and 5FU’s off-target effects and 2) provide critical training to facilitate my transition to independence. My central hypothesis is that obesity induced non- alcoholic fatty liver disease (NAFLD) contributes to disrupted 5FU catabolism and increased toxicity through reduced DPD resulting in reduced functional quality of life and survival. I am proposing a mechanistic aim (1), an exploratory aim (2), and a treatment/intervention aim (3) to test this hypothesis: in aim 1, I will investigate the role of DPD in the metabolism and toxicity of 5FU with obesity (K99); in aim 2, I will explore the impact of 5FU on skeletal muscle and immune cell -omics with obesity (R00); and in aim 3, I will examine the utility of manipulating dietary macronutrients on 5FU’s efficacy and off-target toxicities with obesity (R00). My research aims are complemented by my four training aims: 1) Obesity Phenotyping and Specialized Diet Formation, 2) Plasmid Preparation, CRISPR/Cas 9 Utilization, and –omics, 3) Drug Metabolism (5FU metabolite analysis), and 4) Professional Development and Lab Management. I expect that my findings will provide paradigm shifting evidence for how obese patients should be dosed and monitored to limit chemotherapy’s off-target effects. Additionally, the results from these studies will serve as the foundation for a pathway to independence to continue examining the contributing factors underlying cancer patient life quality and survival. PROJECT NARRATIVE The prevalence of obesity continues to increase in the US and obesity increases the risk of cancer. While it is common practice to apply a dosing cap, current recommendations for the treatment of obese cancer patients is full body surface area chemotherapy dosing regimens despite some evidence suggesting that these patients may have exacerbated drug toxicities. Given that obese individuals are more likely to undergo anti-cancer treatments, there is a need for studying the complex over-lap between obesity and cancer; my proposed study will examine the mechanisms underlying obesity’s impact on the off-target toxicities associated with chemotherapy. | Other Research-Related | 6project_grants_public |
gen_f270b39454ece620b17b10acaf8fb9ac | OCT-based functional biomarkers for degenerative diseases of the photoreceptor-RPE-choroid neurovascular unit | NIH | UNIVERSITY OF CALIFORNIA AT DAVIS | 5R01EY034340-02 | OCT-based functional biomarkers for degenerative diseases of the photoreceptor-RPE-choroid neurovascular unit Abstract: Important diseases of the photoreceptor-RPE-choroid neurovascular unit include age-related macular degeneration (AMD), retinitis pigmentosa (RP), and other inherited retinal degenerations (IRDs). The number of people with AMD worldwide is nearly 200 million, and expected to approach 300 million over the next twenty years, while IRDs affect more than 5 million more. AMD is the leading cause of blindness in the industrialized world, and also the leading cause of blindness among people over the age of 60. Standard treatments exist for neither the more prevalent dry form of AMD nor IRDs, and key obstacles to discovering them are 1) limited understanding of the pathogenic steps leading to vision loss; and 2) lack of biomarkers for disease progression and recovery. In recent years our team has pioneered the emerging field of optoretinography (ORG) using adaptive optics (AO) and OCT. The ORG is an all-optical, noninvasive, objective measure of neural function in the retina, and has the potential to yield new functional biomarkers of retinal disease. Due to the cost and complexity of AO-OCT imaging systems and their subsequent scarcity, ORG measurements have been made only in small numbers of volunteers, mostly without retinal disease. We propose to develop a next-generation, proto-clinical ORG system and characterize its sensitivity, dynamic range, and spatial resolution. First, we will test a number of modifications to the protocols for imaging and signal processing, with a focus on improving the clinical utility of the method. Second, in healthy subjects and several patient populations we will measure ORG responses, along with other OCT-based measurements such as structural OCT imaging and OCTA. The patient populations to be studied are: 1) IRD patients including those with Stargardt’s disease and retinitis pigmentosa; 2) intermediate AMD patients with drusen, subretinal drusenoid deposits, and/or hyperreflective foci; and 3) late AMD patients with regions of geographic atrophy. The motivations for these patient populations are 1) to test hypotheses about the mechanisms underlying the ORG response, and 2) to demonstrate the ORG’s capacity to detect and quantify disease-related dysfunction. The proposed work will result in 1) a clinically useful ORG platform (also capable of structural and angiographic imaging), and 2) new knowledge that will permit us to design ORG- and OCT-based biomarkers for retinal disease that relate to mechanisms of action and have characterized sensitivity to photoreceptor dysfunction. Project Narrative In clinical trials of drugs and other therapies to treat blinding diseases of the retina, the FDA’s recommendation is to use functional assessment, as it is likely to provide earlier indications of their efficacy. In our lab we have discovered a novel way to measure neural function at the cellular level in the living, human retina (the optoretinogram), using equipment very similar to that employed in commercial, clinical imaging systems. The optoretinogram may prove to be faster, less invasive, and more sensitive to neural dysfunction than existing functional tests. The goal of this grant is to study and improve the technology, and then demonstrate its sensitivity to disease-related dysfunction in patients with inherited retinal degenerations and early to intermediate age-related macular degeneration. | Non-SBIR/STTR | 6project_grants_public |
gen_2e4b8a6325c8ced588532d7021e369d5 | Osmotic Regulation of a Peptide Ligand-Mediated Signaling | NIH | RUTGERS THE STATE UNIV OF NJ CAMDEN | 5R35GM151096-02 | Abstract Cells in a multicellular organism constantly communicate by ligand-receptor-mediated signaling to coordinate their growth, development, and adaptation to the environment. The peptide ligands-triggered signaling is the most abundant in this network in humans, dysregulation of which contributes to many prevalent human diseases. However, knowledge of how the peptide ligand-triggered signaling is regulated by the environment remains incomplete. We study a classical peptide ligand-receptor-mediated signaling pathway that regulates the epidermal stomatal development in Arabidopsis thaliana, a naturally simplified and experimentally accessible system. Stomata are micropores on the aerial surface of plants that facilitate gas exchange with the environment. Stomatal development is flexibly adjusted under variable environmental conditions to optimize plant adaptation, making it an attractive system for studies on environmental regulation of peptide ligand-triggered signaling pathways. Upstream of the stomatal signaling pathway are a group of secretory ligands named Epidermal Patterning Factors (EPFs), many of which are highly responsive to environmental cues. We found that osmotic stress caused a dramatic reduction in stomatal density. Interestingly, the transcript of a putative EPF ligand is preferentially induced under osmotic stress. Compared to wild-type plants, the epf mutants produce more stomatal precursor cells under osmotic stress. With these new findings, we hypothesize that osmotic stress inhibits stomatal development via enhancing the candidate EPF ligand-triggered signaling. Our study and others have previously indicated that distinct EPF-triggered signaling pathways target different steps of the stomatal developmental process. The specificity is, at least partially, due to the precise spatiotemporal expression of a particular EPF ligand and the differential subcellular behavior of the receptor kinases including ERECTA-LIKE 1, that transduce the EPF signaling. This MIRA proposal aims to comprehensively evaluate the osmotic stress- induced EPF ligand-triggered signaling pathway by addressing the following three questions: 1) Elucidate the osmotic regulation of the transcription of the candidate EPF gene; 2) Dissect the osmotic regulation of the stomatal receptor complex; 3) Identify the targets of the osmotic stress-induced EPF signaling pathway. Completion of this work is likely to provide valuable insight into our understanding of osmotic regulation of organism development via peptide ligand-triggered signaling pathways. Public Health Relevance Statement Peptide ligand-mediated signaling is integral to the development of multicellular organisms and their adaptation to the environment, dysregulation of which contributes to many prevalent human diseases, including cancer, diabetes, immune-related diseases, and neurological disorders. The proposed study aims at elucidating the osmotic regulation of a classical peptide ligand-mediated signaling pathway. It involves mechanisms of transcriptional regulation, protein subcellular behaviors, protein stability regulation, and cell-state transitioning, all of which are clearly related to the studies of human signaling pathways. | Non-SBIR/STTR | 6project_grants_public |
gen_24a45556acf071ffaf44998779d3180e | Safety and efficacy of Belatacept in heart transplantation | NIH | NEW YORK UNIVERSITY SCHOOL OF MEDICINE | 5U01AI174997-02 | Project Summary: Although calcineurin inhibitors (CNIs), including tacrolimus have led to excellent 1-year outcomes in heart transplantation, long-term survival remains limited by cardiac allograft vasculopathy, driven by donor-specific-antibodies (DSA), and by CNI-associated morbidity, dominated by chronic kidney disease. Belatacept, a selective costimulation blocker, is FDA approved for use in kidney transplantation as a CNI- alternative. By inhibiting CD28/CD80/CD86 interactions belatacept prevents activation of both naïve T cell and T follicular helper cells associated with the development of DSA, but less effectively inhibits memory T cells. In kidney transplant recipients, this has translated to sustained improvement in kidney function, suppression of DSA, and improved graft/patient survival, albeit at the cost of more early rejection which has since been markedly reduced by using a delayed CNI substitution strategy. Building on these findings, we propose a randomized controlled trial of belatacept in first-time heart transplant recipients, in conjunction with gradual tacrolimus withdrawal over 9-months to achieve a CNI-free immunosuppressive regimen (with mycophenolate mofetil and prednisone). We hypothesize that the gradual approach to CNI withdrawal will promote quiescence in graft-reactive T cells, thereby preventing rejection, inhibiting the development of DSA, and eliminating CNI- related morbidity, together increasing survival after heart transplantation. The specific aims are: Aim 1. Clinical trial to determine safety of belatacept in heart transplantation. We will perform a multicenter clinical trial in EBV seropositive heart transplant recipients randomized 2:1 to receive belatacept with gradual tacrolimus withdrawal (9-months) post-heart transplant or standard-of-care tacrolimus (control). The objectives are to a) establish safety of the protocol based on stopping criteria defined by the composite of historical control event rates and b) test efficacy for kidney sparing and DSA. Aim 2. Impact of CNI withdrawal under costimulation blockade on graft-reactive immune responses. We will serially analyze donor reactive, and autoantigen reactive T cell and B cell subsets using state-of-the-art phenotypic and functional assays and will quantify DSA and autoantibodies. Results will be compared between study arms and the kinetics of responses will be evaluated in individual subjects over time. Aim 3. Other mechanistic/biomarker studies relevant to primary and secondary endpoints in the trial. We will use molecular approaches to define differences in the intragraft response, explore markers of kidney injury and fibrosis, and test donor derived cell-free DNA as a potential biomarker of impeding rejection during CNI withdrawal. If successful, belatacept has the potential to transform the heart transplant field, removing CNI-morbidities and preventing DSA as major barriers to improving long-term survival. The comprehensive mechanistic studies will provide novel information, regardless of outcomes of the trial. Project Narrative Long-term survival after heart transplantation continues to be limited both by immune-mediated damage to the transplant and by adverse effects caused by calcineurin inhibitors (CNIs), which are part of nearly all immunosuppressive regimens in heart transplantation. In this application we propose to use belatacept, initially with a CNI that will be slowly reduced and then stopped, to achieve CNI-free immunosuppression (with mycophenolate mofetil and prednisone), which we hypothesize will prevent rejection, suppress the development of antibodies that cause damage to the transplant, and eliminate the adverse effects of CNIs together improving survival after heart transplantation. The accompanying studies will inform as to why the approach did or did not work and determine whether non-invasive biomarkers can be used to predict clinical outcomes. | Non-SBIR/STTR | 6project_grants_public |
gen_e68e887d8f31cc5bb90204841e753068 | Illuminating Lujo virus glycoprotein structure, receptor engagement and neutralizing antibody epitopes | NIH | LA JOLLA INSTITUTE FOR IMMUNOLOGY | 1R21AI180853-01 | Project Summary The Mammarenavirus genus of the Arenavirus family contains multiple zoonotic pathogens with the potential to cause hemorrhagic fever. These include the South American viruses Junin virus (JUNV; Argentinian hemorrhagic fever) and Machupo virus (MACV; Bolivian hemorrhagic fever), and Lassa virus (LASV), which causes thousands of cases of Lassa Fever in West Africa each year. The case fatality rate for these viruses is 20-70%. Lujo virus (LUJV) is the most recently identified African arenavirus. This virus was responsible for five infections, of which four were fatal. Notably, this outbreak was characterized by human-to-human transmission rather than transmission between rodent and human, as is most common for other arenaviruses. Arenaviruses are genetically and geographically divided into New World (e.g. JUNV and MACV) and Old World (e.g. LASV) groups. Its African location placed LUJV into the OW group. However, LUJV is genetically divergent from other African arenaviruses and is phylogenetically equidistant between the NW and OW groups. Further, its glycoprotein GPC recognizes a different receptor and is antigenically distinct from the other arenaviruses. As the only protein expressed on the viral surface, GPC is responsible for receptor engagement, cell tropism and entry, and is the primary target of antibodies. Understanding the unique structure and surface chemistry of LUJV GPC in its native conformation is key to understanding receptor recognition in the native trimer context, what antibody targets might be on this divergent virus, and how we might design vaccines and therapeutics should the virus re-emerge. The premise of this proposal is that structures of the medically relevant LUJV GP alone and in complex with its cell surface receptor will reveal reasons for its unique cell entry requirements and any differences in its epitope landscape. We will use state-of-the-art biophysical techniques such as cryo electron microscopy, surface plasmon resonance and composition-gradient multiangle light scattering, to characterize the interaction of the prefusion-stabilized LUJV GP trimer (pfGP-TD) with its receptor NRP2. In Aim 2, we will identify antibodies from mice immunized with LUJV pfGP-TD using the Berkeley Lights Beacon platform. Results from the innovative research proposed here will launch multiple lines of inquiry for future studies and will help guide development of vaccines against a diverse range of arenaviruses. Project Narrative The arenavirus Lujo virus emerged in Southern Africa in 2008, resulting in a human-to-human transmission chain and deaths of four out of the five infected. Here, we will structurally characterize the surface glycoprotein of Lujo virus alone and in complex with its cellular receptor, and will discover novel antibodies targeting this critical antigen. This work will illuminate the overall architecture and receptor-binding mode of this divergent arenavirus and provide much-needed therapeutic leads and research reagents. | Non-SBIR/STTR | 6project_grants_public |
gen_dcfeb30627a1426e2e99e278ec47a2bc | Mechanisms linking the frail sarcomere to noncompaction cardiomyopathy | NIH | NORTHWESTERN UNIVERSITY AT CHICAGO | 5K99HL168239-02 | Project Summary/Abstract The predominant myosin heavy chain expressed in human heart, beta-MyHC, is encoded by the MYH7 gene. MYH7 variants are well described in hypertrophic cardiomyopathy and less frequently seen in dilated cardiomyopathy. A recent series of publications link variants in the 5’ end of the MYH7 gene as implicated in left ventricular noncompaction cardiomyopathy, often in the setting of a dilated ventricle with impaired function. Importantly, premature truncations as well as missense variation within the MYH7 gene has been linked to LVNC in both population studies and in individuals and families. We now generated a heterozygous premature truncation in MYH7 in human induced pluripotent stem cells (hiPSCs). When differentiated into engineered human heart tissues, we observe the heterozygous premature truncation in MYH7 produces a phenotype consistent with excess proliferation and reduced function, which are key features thought to underlie the development of LVNC in vivo. We hypothesize that truncations and missense variants identified in LVNC are associated with reduced contractility, rather than hyperdynamic MYH7 variants seen in hypertrophic cardiomyopathy. Additionally, many missense variants in MYH7 are considered variants of uncertain significance and methods such as those being used here may help adjudicate variants of risk. Through this training program under the K99 phase, Dr. Monroe will evaluate missense MYH7 variants associated with LVNC and evaluate their performance in engineered heart tissues. In his second aim, he will expand the search for LVNC-associated MYH7 variation to the population scale using linked cardiac imaging and genotype data in the in population datasets. As Dr. Monroe transitions to his independent phase, he will build from work performed earlier in his train implicating the Hippo pathway in proliferation and specification. In Aim 3, he will detail new disease relevance for the Yes-associated protein (YAP) in MYH7-associated LVNC using the models already in hand and further developed under his K99 training. Finally, in Aim 4, Dr. Monroe uses unbiased approaches to characterize human cardiomyocyte heterogeneity in healthy and LVNC engineered heart tissues in order to better delineate the range of differentiation and identify additional downstream pathways that will fuel future investigations. To promote his career development, Dr. Monroe will draw on the strengths of his mentoring committee and primary mentor which will focus on expanding his management and his own mentoring skills. His development plan includes formal and informal courses and workshops aimed at promoting diversity, equity, and research productivity directed towards improving cardiovascular health. Project Narrative Genetic changes in the single gene, MYH7, which encodes the major motor protein in the heart, cause a variety of human heart diseases, called cardiomyopathies. Up to 1/250 individuals carries a genetic change in this gene. Through this work, we will detail why specific genetic defects cause a specific type of developmental heart disease which manifest as cardiomyopathies, and this information will help identify and reduce disease risk. | Other Research-Related | 6project_grants_public |
gen_d79adbe1d804b456013b18415e1b8da8 | Planning & Evaluation | NIH | UNIVERSITY OF CALIFORNIA RIVERSIDE | 5U54CA285114-02 | The goals of the Planning and Evaluation Core (PEC) are to monitor, evaluate, and assess the progress of our U54 Partnerships in meeting its milestones, objectives, and goals. The Core supports our University of California, Riverside (UCR) – City of Hope Comprehensive Cancer Center (CoHCCC) U54 Partnership’s goals of capacity- building, education, and training. Together in this U54, UCR and CoHCCC aim to develop the collaborations, translational resources, and training programs to enhance diversity in cancer research and achieve equity in drug development - throughout the entire drug development pipeline. To achieve our PEC’s goals we are guided by four boards – the Internal Advisory Board (IAB), External Advisory Board (EAB), Community Advisory Board (CAB), and Diversity, Equity, and Inclusion Board (DEI). Our IAB and EAB have served a key role in guiding our P20 planning grant; in our proposed U54 these two boards will be joined by our newly created CAB and DEI. The inclusion of these two new boards underscores our U54’s commitment to 1) serving the diverse individuals that live in our Southern California communities and 2) to mentor and train a diverse early-stage investigators who are committed to eliminating our current disparities in cancer therapeutics, drug development, and access to clinical trials. Through the activities and oversight of the PEC, our four advisory boards will 1) conduct evaluation, tracking, and reporting of the progress of projects and cores and 2) provide focused feedback to program U54 leadership and investigators. As in our P20 planning grant, the U54 PEC will work with the IAB to provide a structured and rigorous review process to select new Full and Pilot projects as well as Mini-Pilots for the Capacity Core. The PEC will assess all components of our U54 partnership. The PEC will assure that the 1) Research and Evaluation Core training and education programs are effective in providing education and building capacity, 2) Outreach Core works with investigators to be inclusive of the communities we serve, and 3) Capacity Core identifies provides early-stage investigators (ESI) with mentorship in target identification, drug development, and clinical trials. We anticipate that together our Cores will play a key role in developing future U54 projects that build UCR’s capacity in drug development and clinical trials. | Research Centers | 6project_grants_public |
gen_70596c6ee3d872d2723b89c1cec1d03b | Synthesis of Bioactive Natural Products and Unnatural Congeners | NIH | FLORIDA STATE UNIVERSITY | 5R35GM150795-02 | Project Summary/Abstract Alkaloid natural products have had an immense impact on the field of therapeutic medicine, and hold promise for the discovery of new neuropharmacological treatments. One area of proposed research in the Smith Lab concerns the development of new psychoplastogenic compounds based on the psychedelic lysergic acid diethylamide (LSD). Although derivatives of LSD have been explored previously over many decades, derivatives bearing various substitutions on the indole ring of the ergoline scaffold have never been explored. All of these derivatives are currently only accessible through total synthesis, and potentially can serve as therapeutic leads for various neuropsychiatric diseases including Parkinson's disease, PTSD, severe depression, and addiction. Our concise synthetic approach to the scaffold of LSD hinges on the utilization and chemical modification of widely available heterocyclic aromatic precursors. This strategy allows for the exploration of chemical space that would otherwise be inaccessible, however therapeutically enabling. Synthetic inspiration drawn from our route to these LSD analogs has led us to investigate the synthesis of marine macrocyclic diamine alkaloids such as the halicyclamines and the sarains. None of these targets have been synthesized previously. Our modular, yet redox-economic platform for assembling these structurally complex compounds hinges on employing heterocyclic starting materials at higher oxidation levels to efficiently access their congeners with lower redox levels. Innovation in this research direction is mainly strategic, demanding a navigation of synthetic assembly that has not been traversed previously. This will also demand tactical advances in chemistry that will undoubtedly shed light on novel reactivity patterns that will be broadly important within the realm of heterocyclic chemistry. Project Narrative Alkaloid natural products have had an immense impact on the field of therapeutic medicine, and hold promise for the discovery of new neuropharmacological treatments. The development of new synthetic strategies to build these important molecules allows for the construction of new matter capable of improving upon the available treatments for crippling ailments such as addiction, depression, post-traumatic stress disorder and Parkinson's. Laboratory synthetic access to these important alkaloid natural products holds immense promise for the development of efficacious and novel neuropharmacological small molecules. | Non-SBIR/STTR | 6project_grants_public |
gen_7091e600bc4e516a5fe1dda6de50c976 | Reducing Stigma in People Who Inject Drugs with HIV Using a Rapid Start Antiretroviral Therapy Intervention | NIH | YALE UNIVERSITY | 3R01TW012674-02S1 | HIV transmission continues in low- and middle-income countries (LMIC), especially among key affected populations (KAP) and in settings of high stigma and discrimination. In Malaysia, a LMIC in SE Asia, HIV incidence and mortality is increasing. HIV is concentrated among KAPs, especially people who inject drugs (PWID), a group that has substantially lower ART prescription and viral suppression (VS) levels relative to other KAPs, undermining HIV treatment as prevention (TasP) goals. PWID are especially vulnerable to overlapping and intersectional stigmas due to criminalization drug use and sex work, experiences with incarceration, social class and the presence of HIV itself. Our preliminary studies confirm high levels of negative stereotypes, prejudice and stigma toward PWID among medical students and HIV experts, with clear evidence of intention to discriminate against PWID by withholding ART prescription. Stigma-reducing interventions have mostly centered on educational and contact-based strategies. Such strategies, however, appear less effective where stereotypes and stigma are deeply entrenched, as in Malaysia, thus requiring the introduction and testing of alternative strategies. Behavioral design interventions are potentially effective ways to address stigma in such settings. Behavioral design interventions use tools like framing, nudges, and choice architecture, which can be used to re-design how physicians behave – or make non-discriminatory healthcare decisions. Rapid start antiretroviral (RS-ART) is an evidence-based strategy to initiate ART immediately, thereby supporting TasP goals by reducing time to VS, achieving VS and improving individual health. It has not been tested among PWID. It fits the criteria for behavioral design interventions by re-arranging clinician decision-making by first focusing on eligibility criteria (i.e., presence of opportunistic infections) rather than inaccurate perceptions of ART adherence or deservedness. Behavioral design interventions have not been tested in HIV stigma research, nor has they been assessed longitudinally or infusing clinically relevant dyads analyses of patients and clinicians. To guide the behavioral design of RS-ART among PWID, we will use the Delphi method to develop guidelines. Then we will use nominal group technique, a rank-ordering mixed method strategy to assess the multi-level barriers and facilitators to RS-ART for PWID, in order to adapt existing RS-ART protocols for PWID. Once the new guideline concordant RS-ART protocol is developed, we will pilot test it in 125 PWID over six months and conduct a longitudinal dyadic analysis of patients and clinicians of stigma, physician trust and social support. The RS-ART protocol will be refined further during pilot-testing to determine its utility as a stigma-reducing intervention that can be tested in a future implementation trial. This proposal brings over 17 years of productive collaboration between Yale and University of Malaya, with expertise in clinical HIV and addiction treatment, participation in clinical guidelines development, mixed methods research, intervention development and refinement, multi-level stigma assessment and intervention and dyadic analyses. Malaysia, a middle-income country that has an expanding HIV epidemic concentrated in people who inject drugs is steeped in high levels of discrimination and intentions to discriminate by HIV treating clinicians. We propose to develop and pilot test a rapid start antiretroviral therapy strategy that we will pilot test as a stigma reduction intervention, which if effective, can be tested in future implementation trials. | Non-SBIR/STTR | 6project_grants_public |
gen_8222ea247a3f176ebc989b6716bde3ed | Effect of weight loss on intermuscular adipose tissue (IMAT) signaling | NIH | UNIVERSITY OF COLORADO DENVER | 3R01DK134706-02S1 | Project Summary/Abstract Black Americans experience disproportionality high rates of obesity, insulin resistance and diabetes and may derive fewer metabolic benefits from weight loss compared to their White counterparts. The mechanisms responsible for these differences are poorly understood as Black men and women are typically underrecruited in clinical studies. Intermuscular adipose tissue (IMAT) is marbled within and next to skeletal muscle and can cause muscle insulin resistance by bathing the muscles in inflammatory cytokines, adipokines, eicosanoids, and free fatty acids (the IMAT secretome). The literature shows that Black individuals have greater IMAT content and adipose tissue inflammation compared to White individuals. Preliminary data show that IMAT has a similar or worse secretome as visceral adipose tissue towards muscle insulin sensitivity, with secretion of inflammatory cytokines and eicosanoids contributing to muscle insulin resistance. Our central hypothesis is that IMAT secretion of inflammatory cytokines is greater in Black than White participants with less improvement following weight loss, contributing to the racial differences in insulin sensitivity. The purposed research addresses the critical need to elucidate disease mechanisms in Black individuals by comparing changes in the IMAT secretome before and after weight loss between Black and White participants. The rationale is that we will generate new information for differences by race in IMAT paracrine signaling and generate new information for changes in the IMAT secretome by race after weight loss. By identifying differences between obese Black and White participants in the IMAT secretome before and after weight loss, this critical research can support development of tailored diabetes interventions and treatments. We propose two specific aims: Aim1: Evaluate the potency of IMAT paracrine signaling to decrease insulin sensitivity in Black and White participants. We hypothesize IMAT secretes greater inflammatory cytokines and eicosanoids linked to decreased insulin sensitivity in Black compared to White participants. In vitro experiments will measure the potency of the IMAT secretome to cause insulin resistance by race. Aim 2: Determine the effect of weight loss on the IMAT content and paracrine signaling in Black versus White participants. We again hypothesize that weight loss will diminish IMAT secretion of inflammatory cytokines and eicosanoids linked to decreased insulin sensitivity, but this decrease will be less in Black compared to White individuals. Participants with obesity will be studied before and after a 12-week weight loss intervention. IMAT will be sampled using an ultrasound-guided IMAT biopsy technique, insulin sensitivity measured using hyperinsulinemic-euglycemic clamps, and IMAT content measured using MRI. IMAT will be cultured to generate conditioned media, followed by conditioned media analyses and testing of its direct metabolic effects in vitro. This project will reveal racial differences in IMAT secretome, and malleability after weight loss that will inform tailored development of interventions to prevent IMAT-induced insulin resistance. . Project Narrative: Relevance of research to public health This project will investigate metabolic differences in the intermuscular adipose tissue (IMAT) secretome between obese Black and White participants before and after weight loss. Moreover, it will employ culturally sensitive strategies to recruit and retain Black participants. These findings are crucial for developing diabetes interventions and treatments tailored specifically for the Black population. | Non-SBIR/STTR | 6project_grants_public |
gen_b0edafb6a3fececbe00700122d67afe2 | Actin regulation by cyclase-associated protein | NIH | EMORY UNIVERSITY | 1R01GM144563-01A1 | Project Summary Reorganization of the actin cytoskeleton supports many dynamic cell biological processes, such as cell migration, cytokinesis, vesicle trafficking, and morphogenesis. Although actin spontaneously polymerizes and depolymerizes by itself, the dynamic properties of actin need to be controlled by actin-regulatory proteins in cells. Among many known actin-regulatory proteins, cyclase-associated protein (CAP) regulates multiple key processes of actin dynamics. CAP is conserved among eukaryotes and has been known as an actin-monomer binding protein for many years. However, recent studies have identified important functions of CAP, which are critical to promote rapid actin turnover. CAP promotes disassembly of actin filaments from the pointed ends in the presence of actin-depolymerizing factor (ADF)/cofilin. Spontaneous actin depolymerization from the pointed ends is very slow and rate-limiting in actin filament turnover. The synergy between CAP and ADF/cofilin can enhance actin depolymerization in vitro to a range that can explain rapid actin turnover in cells. However, the precise mechanism by which CAP and ADF/cofilin induce actin depolymerization remains unclear, partly because structural information of a CAP oligomer is limited. CAP oligomerizes under physiological conditions, and oligomerization enhances its activity. We have recently obtained evidence that CAP is tetrameric, and our structural model of the CAP tetramer allowed us to hypothesize a functional link between the tetrameric configuration of CAP and processive actin depolymerizing activity. CAP also binds to the side of actin filaments and synergizes with ADF/cofilin to promote filament severing. We hypothesize that this is due to the effect of CAP on the helical structures of actin filaments, which then causes partial dissociation of ADF/cofilin from the filaments. In addition to the in vitro studies, recent reports of human mutations in the CAP (CAP2) gene have linked impairment of CAP to human diseases in the heart and skeletal muscle involving abnormalities in the actin cytoskeletal organization. We will use the nematode Caenorhabditis elegans as a relevant model organism to study roles of CAP in the assembly of sarcomeric actin filaments in muscle cells in vivo. We proposes three aims: (1) to determine functional significance of CAP tetramerization for actin filament disassembly, (2) to determine the effects of CAP on the helical structures of F-actin, and (3) to determine in vivo roles of CAP in actin cytoskeletal integrity in the nematode Caenorhabditis elegans. We will employ a combination of in vitro and in vivo approaches to gain new insight in the function of CAP in actin cytoskeletal regulation. Project Narrative Dynamic reorganization of the actin cytoskeleton is essential for cell migration, cell division and morphogenesis. The basic regulatory mechanism of actin dynamics will be investigated by a combination of biochemical and genetic approaches. | Non-SBIR/STTR | 6project_grants_public |
gen_ffec76512ebcbb1a99d5efdea51d128e | Development of Potent Estrogen Receptor Beta Agonists for Treating Glioblastoma | NIH | UNIVERSITY OF TEXAS HLTH SCIENCE CENTER | 1R01CA269866-01A1 | Glioblastoma (GBM) is the most common primary malignant brain tumor with a survival time of approximately 19 months and the 5-year survival rate is ~10%. Standard treatment for GBM consists of surgical resection, external beam radiation therapy (XRT), and adjuvant chemotherapy with temozolomide; however, resistance to XRT and chemotherapy is a major clinical problem. Recent studies suggest female sex hormones play a protective role in GBM progression. However, the utility of using estrogen as a treatment for GBM is limited due to its associated toxicity and risks of developing new cancers. Estrogen functions are mediated by two estrogen receptor (ER)- subtypes: ERα that functions as a tumor promoter and ERβ that functions as a tumor suppressor. Recent studies using CRISPR KO in human GBM models have confirmed that ERβ functions as a tumor suppressor in GBM. Nonetheless, the therapeutic potential of ERβ have not been extensively exploited. Currently available synthetic ERβ agonists (LY and ERB041) are proven to be safe for human use; however, these are no longer in clinical development by industry due to failure to meet clinical endpoints in non-oncological clinical studies. Low efficacy of synthetic ERβ agonists is ascribed in part to requiring high concentrations (10-100 µM) resulting in their cross reactivity with ERα. Therefore, the development of novel selective ERβ agonists, with higher selectivity and high potency is needed for clinical translation. In collaboration with the Center for Innovative Drug Discovery (CIDD) at UTSA, we have developed lead ERβ agonists that deliver higher potency and specificity to ERβ, which we have branded as CIDD-ERβ agonists. The objective of this proposal to translate the functional role of ERβ as a tumor suppressor into a clinical strategy utilizing novel CIDD-ERβ agonists as a new therapeutic agent. The hypothesis is that potent ERβ specific agonists block GBM progression by promoting growth inhibitory pathways and sensitizes them to radiation and chemotherapy. We will test this hypothesis using three aims. In Aim 1, we will further optimize the translatability of CIDD-ERβ agonist leads by using its structure- based design, medicinal chemistry approaches and develop CIDD-ERβ agonists with higher specificity, potency and central nervous system (CNS) ADME properties. Further, we will determine maximum tolerated dose, toxicology and establish PK, PD. In Aim2, we will confirm the specificity of interaction of CIDD-ERβ agonists with ERβ using biophysical methods and confirm the effect of CIDD-ERβ agonists on ERβ genomic, non-genomic and DNA damage response functions. In Aim3, we will test the efficacy of optimized CIDD-ERβ agonists on glioma stem cells (GSCs), test their efficacy on tumor progression and survival using patient xenograft GBM models and also test the efficacy in conjunction with radiation and chemotherapies. This proposal is clinically significant as successful testing of these hypotheses will result in the development of novel ERβ agonists that promote tumor suppression, which can be readily translated into clinical use simultaneously with current chemo and radiation therapies, providing an additional tool for enhancing survival in GBM patients. Glioblastoma (GBM) is the most common primary malignant brain tumor in adults with poor survival. This research proposal is based on strong preliminary data showing that Estrogen Receptor beta (ERβ) exerts tumor suppressive functions in GBM. This proposal is will develop novel ERβ agonists that promote tumor suppression leading to a new therapeutic modality to treat GBM. Identification of CIDD-ERβ agonists as a therapeutic agent can be readily translated into clinical use with current chemo and radiation therapies, providing an additional tool for enhancing survival in GBM patients. | Non-SBIR/STTR | 6project_grants_public |
gen_e775f2347baa82d606714a69f8a157ce | Genetic interactions among targets of master regulator genes as drivers of complex behavior in Drosophila intestinal stem cells | NIH | CALIFORNIA STATE UNIVERSITY NORTHRIDGE | 1R16GM149452-01 | PROJECT SUMMARY Several master regulator (MR) genes have been characterized in tissue stem cells across organs and species. However, they remain poor candidates for therapeutic manipulation because they are highly pleiotropic, affecting hundreds of targets and operating across many organ systems. Therefore, only a better understanding of how less pleiotropic downstream MR targets coordinate stem cell proliferation, self-renewal and differentiation will unlock the full potential of stem cells in regenerative medicine. The long-term goal of this project is to map the regulatory landscape established by MR genes and their targets in intestinal stem cells (ISCs), using the fruit fly Drosophila melanogaster as a model system. Drosophila ISCs divide asymmetrically, giving rise to a new ISC and a sister that will become an absorptive enterocyte (EC) or a hormone-secreting enteroendocrine cell (EE). The specific hypothesis driving this proposal is that cross-regulatory interactions between MR target pathways can lead to non- linear, unpredictable outcomes on ISC behavior when they are manipulated simultaneously. To test this hypothesis, CAP, Klaroid and Indy, three experimentally validated targets of the ISC MR genes Escargot and STAT, will be manipulated alone or in combination within ISCs using an inducible Gal4/UAS system. The effect of their individual vs. combined manipulations on intestinal homeostasis will be assessed via three separate but complementary approaches. In Aim 1, immunofluorescence microscopy will be used to compare ISC number, morphology, mitotic rate, and differentiation potential, based on well-established cell type markers (esg-GFP for ISCs, Su(H) activation for EBs, Pdm1 and Pros staining for ECs and EEs, respectively). Automated image analysis through ImageJ and CellProfiler will be used to analyze multiple images per group, allowing a robust statistical analysis of the data. In Aim 2, fluorescent activity reporters will be used to compare the effect of single vs. dual MR target manipulations on key ISC signal transduction pathways (EGFR, Notch, Wnt, STAT and JNK). In Aim 3, lifespan and intestinal barrier integrity (Smurf) assays will be used to compare the effect that individual vs. combined MR target manipulations have on the regenerative capacity of intestinal tissue following chemical injury or pathogenic infection. These research aims may generate evidence that challenges the widely held premise that combination therapies can only improve outcome due to additive complementation of positive effects. If so, this project will have a significant impact on our conceptual approach to stem cell manipulation for regenerative medicine. In addition, this project was specifically designed to engage a large number of students from underrepresented backgrounds in biomedical research, satisfying another important mission of the NIH: to diversify the scientific workforce, and thus foster innovation, improve research quality and enhance the public trust and investment in science. PROJECT NARRATIVE Although decades of research have identified genetic master regulator (MR) genes that are essential for the regenerative function of stem cells, the downstream mechanisms that they regulate remain largely unknown. This project seeks to characterize the function of a small set of validated MR targets in the regulation of intestinal stem cells, using the fruit fly Drosophila melanogaster as an experimental model system. The research proposed is dually relevant to the NIH mission, for it will contribute to our basic understanding of how stem cells can be used safely and effectively in regenerative medicine, while specifically expanding the repertoire of therapeutic targets for the treatment of gastrointestinal disease. | Non-SBIR/STTR | 6project_grants_public |
gen_39a1084edecce6faa687c722dabc4a8d | Leveraging Health Systems to Increase Implementation of Evidence-based Surgical Cancer Care | NIH | UNIVERSITY OF ALABAMA AT BIRMINGHAM | 5K08CA283001-02 | PROJECT SUMMARY Hospitals are increasingly consolidating into health systems with shared ownership and management. Care in a health system has potential benefits for surgical cancer patients including improved access, care coordination, and strategies to disseminate and implement a rapidly evolving evidence-base into practice across system hub and spoke sites. Prior research has demonstrated that these potential benefits remain elusive. The impact of consolidation on quality varies widely, and there are disparate outcomes for surgical cancer patients treated at different facilities in the same systems. We have shown that location of surgical cancer care determines whether effective treatments are adopted (implementation) or ineffective treatments discontinued (de- implementation). We hypothesize that health system characteristics and strategies are associated with variability in implementation of oncologic evidence among hub and spoke hospitals and that through exploration of observed differences we will identify levers for targeted, multi-level interventions. This work addresses the NIH Blueprint objective to enhance research investments by ensuring adoption into practice and targets rural individuals, a population with disparate healthcare access and outcomes, who are often treated at spoke hospitals. We will examine the influence of treatment for common cancers in health system hubs and spokes on patient access and receipt of evidence-based care by linking SEER-Medicare data with health system data. Then, we will identify health system characteristics associated with evidence implementation, both quantitatively using multilevel modeling and qualitatively through structured interviews with health system stakeholders. Finally, we will use the resources within our Health System at the University of Alabama at Birmingham (UAB) to develop a system-level intervention for dissemination and implementation of oncologic evidence across hub and spoke sites. My long-term goal is to become an independent investigator who improves the quality of cancer care delivery by designing, implementing and studying health system-level interventions to increase clinical adoption of oncologic evidence. Through this training award, I will complement my health services and quality improvement science expertise with advanced training in the organization of healthcare delivery, multilevel analysis of secondary data, and implementation science to develop a system-level intervention to improve evidence-based surgical cancer care. PROJECT NARRATIVE Most patients with common cancers in the United States receive surgical care within health systems, but quality is inconsistent across system hub and spoke sites. This project will evaluate the effect of treatment in a health system hub or spoke on receipt of evidence-based surgical cancer care and will identify health system characteristics and strategies that are associated with system performance and consistency across sites. The product, an intervention map for system-level implementation of oncologic evidence, can be used by health systems to provide consistent, high-quality care across sites, increasing equity in cancer care delivery. | Other Research-Related | 6project_grants_public |
gen_db62ac329e9c9183324b0d4585b7662d | High-throughput, purification-free, and ultrasensitive transmembrane nanosensor arrays for digital counting of microRNA biomarkers of intact exosomes | NIH | ARIZONA STATE UNIVERSITY-TEMPE CAMPUS | 5R61CA278558-02 | Project Summary/Abstract Rapid and specific histopathologic diagnoses are critical for cancer treatment. Tumor tissue biopsy is routinely performed to detect and monitor cancer progression. Current test biopsies require surgically-collected tissue samples from detectable primary or metastatic tumors. Several difficulties, such as patient inconvenience, multistep complicated procedure, partial samplings, and non-specific findings, make this process slow, invasive, expensive, unfit for screening large sample sizes, and error-prone. Non-invasive selections of biomarkers in body fluids, known as liquid biopsy, offer great promise in complementing or even substituting surgical tissue biopsy in the diagnosis and prognosis of cancer patients. Recent studies have indicated exosomal microRNAs (exmiRs) as promising liquid biopsy biomarkers in detecting cancer progression and efficacy of therapy with high sensitivity and specificity. However, current technologies for ex-miR detection, such as qRT-PCR, and microarray screening tests, require high sample volume, are expensive, slow, tedious, requiring highly specialized skills and resources such as ultracentrifuge, expensive RNA extraction kits, etc. Single-exosome level studies can significantly circumvent these problems. However, the few single-molecule ex-miR quantification attempts lack amplification strategy, thus limiting their applications to resource-heavy research settings. To address these problems, we have developed a molecular beacon-based Transmembrane Nano-Sensor (TraNS) that inserts itself into the membrane of lipid vesicles and signals the presence of a DNA target by an increase in fluorescence. We have successfully demonstrated the ability of the TraNS device to spontaneously insert into the lipid membrane and sense membrane-enclosed nucleic acid biomarkers with high specificity. In this study, we propose to (1) optimize the TraNS device to sense cancer- specific ex-miRs from biofluids, (2) harness the transmembrane structural reconfiguration of TraNS to develop an isothermal signal amplification method to improve the sensitivity of detection significantly, and (3) integrate the TraNS device with our patented DNA origami-based biomarker detection array to improve the throughput, specificity, and sensitivity of digital quantification of ex-miR stoichiometry with low sample volume. We will use the platform’s sensitivity, specificity, and throughput on clinical samples from pancreatic cancer patients against their healthy controls. This effort’s potential impact can help physicians and clinicians with rapid, ultrasensitive, precise, and cost-effective cancer diagnostics without a surgical tissue biopsy. Project Narrative/Relevance We propose to develop a transmembrane sensor nanoarray platform that, if successful, will allow rapid, ultrasensitive, and specific detection and quantification of exosomal micro-RNA at the level of single exosomes. We will design Aampanying isothermal signal amplification strategies to detect the low abundance of nucleic acid biomarkers in exosomes. If successful, the proposed work will provide scientists, physicians, and clinicians with a means for rapid, ultrasensitive, highly-specific, cost-effective prognosis and diagnosis of cancer patients without surgical tissue biopsies. | Non-SBIR/STTR | 6project_grants_public |
gen_d1487f6876d85b7ac11c6cb4d866794f | Development of novel malaria pre-erythrocytic vaccine antigens targeting Plasmodium sporozoite liver infection | NIH | MERCER UNIVERSITY MACON | 5R21AI166451-03 | Previously, using a phage peptide display library, our group has identified Plasmodium parasite ligands and corresponding host cell receptors important for sporozoite-Kupffer cell interaction in the mammalian liver. Using a similar approach, we identified the HP1 peptide, a structural mimic of a ligand that the sporozoite uses to infect hepatocytes. Immunization with the HP1 peptide protects ~ 50 % mice from P. berghei challenge. Using an anti-HP1 antibody, we identified Plasmodium Phospholipid Scramblase (PLS) as the sporozoite ligand of hepatocytes and a potential vaccine antigen. We propose 1) to identify PLS protein epitopes that function in hepatocyte recognition, for use as a vaccine antigen; and 2) to develop a tri- functional vaccine antigen containing i) a sporozoite circumsporozoite protein (CSP) epitope, targeting liver sinusoid binding, ii) a sporozoite GAPDH-related epitope, targeting sporozoite exit from the circulation via Kupffer cell traversal, and iii) sporozoite PLS epitope, targeting hepatocyte infection. We expect that this approach will lead to enhancement of the moderate protective efficacy of the most advanced RTS,S/AS01 malaria vaccine. Malaria is among the deadliest infectious diseases and kills an estimated half million persons every year. This project seeks to identify vaccine epitopes that target three critical stages of parasite infection of the liver. The findings may lead to the development of a novel and improved malaria pre-erythrocytic vaccine. | Non-SBIR/STTR | 6project_grants_public |
gen_9a3c2b22857cadfcf08478557d371e5d | Sorting and characterization of mechanically heterogeneous cell populations based on cellular contractility | NIH | UNIVERSITY OF ARKANSAS AT FAYETTEVILLE | 1R21EB034930-01 | Project Summary Cellular contractility plays a critical role in both development and disease. Recent evidence suggests that even within a cell population derived from the same source, vast heterogeneity exists in terms of cellular contractility. Dysregulation of spatiotemporally organized cellular contractility often results in developmental defects. In invasive diseases like cancer which are often highly contractile, the existence of a weakly contractile subpopulation is receiving increasing attention. Adherent cells are known for their ability to sense and dynamically adapt to their local microenvironment. Hence, the heterogeneity in mechanical phenotype may be a result of genetic heterogeneity or cellular plasticity and mechanical adaptation. Mechanomedicines or mechano- based therapies that target specific physical cellular and tissue interactions, including abnormal cellular contractility, in diseases like cancer, fibrosis, and cardiovascular disease, as well as aging, are emerging and hold great potential. While mechanical heterogeneity and plasticity are known to contribute to resistance to therapies that target a specific molecular pathway, it is not clear whether a change in mechanical phenotype predicts disease outcome or if mechanical adaption happens as a result of disease progression. This project proposes to phenotypically sort adherent cells into subpopulations with distinct contractile phenotypes and use these sorted subpopulations to test the hypothesis that the initial contractile phenotype and heterogeneity determine the disease outcome against the alternative hypothesis that mechanical adaptation to the local microenvironment and phenotypical switching contribute to disease progression regardless of the initial mechanical heterogeneity. Cancer metastasis will be used as the main biological model for hypothesis testing. In Aim 1, the engineering approach for cell sorting based on cellular contractility will be optimized. Fluorescence- activated cell sorting (FACS) will be coupled with an engineered high throughput cell contractility screening platform, automated microscopy, and photoactivation and fluorescent labeling of cells for cell separation. In Aim 2, the sorted contractile subpopulations will be used to test the main biological hypothesis in vitro and in vivo against the alternative hypothesis. Engineered systems mimicking the environmental conditions in cancer progression will be designed to characterize the migration, proliferation, survival, and metabolism of the subpopulations, as well as their mechanical adaptation. The metastatic potential of these subpopulations and their mechanical adaptations at various stages along the metastatic cascade will be examined in a mouse tumor model. The innovative aspect of this proposal is the concept to sort by cellular contractility with the goal of uncovering the role of initial mechanical phenotype in the progression of diseases like cancer and in development. This project will use the novel engineered cell sorting approach to dissect the respective roles of mechanical heterogeneity and adaptability in disease progression, thus laying the foundation for future work to identify the key molecular pathways to precisely target for the development of mechanomedicine. Project Narrative Cellular mechanical properties such as contractility play key roles in development and are often altered in diseases, thus targeting the altered mechanical properties of cells holds great potential. However, diseased cells often exhibit different mechanical phenotypes and can switch between these different phenotypes during disease progression, which allows them to resist and escape therapeutic targeting. This project develops an engineered high-throughput platform to isolate and characterize mechanically distinct cell subgroups based on cellular contractility, thus providing a versatile tool to understand the effect of distinct mechanical phenotypes on the progression of human diseases such as cancer. | Non-SBIR/STTR | 6project_grants_public |
gen_229b73c8a244e4a134bf894e9663619e | Mechanotransduction via LIM Domain Protein Mechanosensing | NIH | LOYOLA UNIVERSITY CHICAGO | 5R01GM148644-02 | Project Summary Mechanical interactions play a fundamental role in physiology, allowing cells to move, generate forces, and assemble into multicellular structures. Key to these processes is the ability of cells to turn mechanical signals into biochemical signals, an activity known as mechanotransduction. The search for mechanosensitive proteins that could facilitate mechanotransduction has primarily focused on proteins that undergo conformational changes in response to force or proteins that display changes in the bond kinetics under load. There exists another class of proteins, however, that recognize structures under strain. The canonical example of this class of proteins is the LIM domain protein zyxin, which recognizes strained actin stress fibers and recruits actin polymerization factors to repair them. Recent work has highlighted that the strain sensing mechanism of the LIM domains is not unique to zyxin and that numerous other members of the family of LIM domain proteins display a similar ability. This suggests LIM domain proteins could act as mechanotransducers, recognizing strain via their LIM domains and converting it to other biochemical signals via interactions with the other domains in the protein. To explore this hypothesis further it is crucial that we understand how LIM domains recognize strained actin filaments, and how those interactions propagate signals downstream of the strain sites. Here we propose to establish rigorous experimental strategies to decipher the mechanisms underlying LIM domain protein mechanotransduction. We employ a combination of biophysical techniques including laser ablations, optogenetics and cell stretching to quantitatively and repeatedly induce strain sites in the actin cytoskeleton. In Aim 1 we will test alternative mechanisms of LIM domain strain sensing by comparing proteins from the testin family of LIM domain proteins which require only a single LIM domain to recognize strain sites, compared to the three tandem LIM domains that zyxin requires. In Aim 2 we will test whether in addition to stretched actin filaments in stress fibers, LIM domain proteins can recognize other strained actin structures, such as compressed stress fibers or actin meshworks. Finally, in Aim 3 we will investigate how binding of LIM domain proteins to strain sites leads to a propagation of that mechanical signal to other parts of the stress fiber and the extracellular matrix. Together these studies will greatly expand our knowledge of mechanotransduction and provide insight into this fundamental signaling mechanism. Project Narrative Mechanical interactions and signals provide the basis for numerous physiological processes including allowing cells to move and form multicellular structures. Misregulation of these mechanical signals has been implicated in a broad array of diseases and developmental impairments. This project investigates novel mechanisms for how proteins can sense these mechanical signals and convert them to biochemical signals, thereby uncovering new signaling pathways and potentially identifying new therapeutic targets. | Non-SBIR/STTR | 6project_grants_public |
gen_82663bbb4d9a3d0fef1f374482bb75ab | Explant human skin perfusion model to study mechanisms of chemical injury and mitigation | NIH | UNIVERSITY OF PITTSBURGH AT PITTSBURGH | 1R34AR083572-01 | The changing geopolitical environment has increased the risk of mass population exposure to chemical vesicants-induced skin injuries from a large-scale incident improvised by a state or terrorist organization executed act but there are few countermeasures available having limited efficacy. The long-term goal is to develop a potent therapy for the treatment of chemical vesicant-induced injuries and its addition to the national stockpile. The overall objectives of this proposal are to (i) equip our laboratory with trained human resources and facilities that enable us to conduct research in the area of chemical vesicant-countermeasures development and (ii) understand the dynamics and mechanism(s) of vesicants-induced skin injuries. The central hypothesis is that understanding the dynamics of human skin response and the mechanism of the response to chemical vesicants is the key to designing efficient countermeasures. The rationale for this project is that our novel human skin perfusion model offers a unique opportunity to test the dynamics and mechanism(s) of human skin injury to vesicants in a controlled environment. The central hypothesis will be tested by pursuing three specific aims: 1); Equip our laboratory and acquire essential training to handle toxic vesicants and human skin perfusion system. 2); Implement a human skin bioreactor workflow for rigorous evaluation of the effects of NM and PAO vesicants 3) Investigate the mechanism(s) of vesicant-induced cell injury and death pathways in human skin. Under the first aim, we will procure additional safety equipment and get the training to handle toxic vesicants and operate the human skin perfusion system. The second aim will focus to study the dynamics of vesicants-induced injury in response to different doses and exposure times. While the third aim is focused to determine the mechanism(s) of the injury. The proposal is innovative, as it aims to perform pre-clinical mechanistic studies using a novel platform in human tissues. The proposed research is significant because the successful completion of this project will provide a strong scientific platform for investigating targeted countermeasures strategies against vesicant-induced skin injuries and due to the use of the human tissue as a model the approval pathway of countermeasures for inclusion into the national stockpile can be accelerated. Project Narrative The proposed research is relevant to public health because it will facilitate and vertically advance the development of critically needed medical countermeasures against chemicals induced skin injuries. The proposed mechanistic studies will facilitate understanding the dynamics of the key processes involved in the chemicals induced injuries that will have an enormous impact on our capabilities to design countermeasure strategies and test them directly on human skin using our novel model system. This whole process will shorten the time for developing countermeasure and increase the chances of success in clinical translation Thus, the proposed research is relevant to the NIH/NIAID/Chemicals Countermeasure Program’s mission to develop new countermeasures and add them to the national stockpile. | Non-SBIR/STTR | 6project_grants_public |
gen_735b54f3ec7b01bb704160d10b2f4c7e | LEAarning analytics and AI for personaliseD lEaRning | European Commission (EC) | VIRTUALCARE, LDA;Πανεπιστήμιο Κρήτης – Τμήμα Βιολογίας;CENTRE FOR ADVANCEMENT OF RESEARCH AND DEVELOPMENT IN EDUCATIONAL TECHNOLOGY LTD-CARDET;EDEX - EDUCATIONAL EXCELLENCE CORPORATION LIMITED;Tallinn University;University of Pitesti | Not available | Unknown | 6project_grants_public |
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